void pulsesequence() { /* DECLARE & READ IN NEW PARAMETERS */ char compshape[MAXSTR]; getstr("compshape",compshape); /* Composit pulse shape */ loadtable("lc1d"); /* Phase table */ /* PULSE SEQUENCE */ status(A); hsdelay(d1); status(B); if (getflag("wet")) wet4(t1,t2); status(C); if (getflag("composit")) { if (rfwg[OBSch-1] == 'y') shaped_pulse(compshape,4.0*pw+0.8e-6,t3,rof1,rof2); else composite_pulse(pw,t3,rof1,rof2,v1); } else rgpulse(pw,t3,rof1,rof2); setreceiver(t4); }

pulsesequence() { // Define Variables and Objects and Get Parameter Values MPSEQ dec = getblew("blewH",0,0.0,0.0,0,1); strncpy(dec.ch,"dec",3); putCmd("chHblew='dec'\n"); CP hx = getcp("HX",0.0,0.0,0,1); strncpy(hx.fr,"dec",3); strncpy(hx.to,"obs",3); putCmd("frHX='dec'\n"); putCmd("toHX='obs'\n"); //-------------------------------------- // Copy Current Parameters to Processed //------------------------------------- putCmd("groupcopy('current','processed','acquisition')"); // Dutycycle Protection DUTY d = init_dutycycle(); d.dutyon = getval("pwH90") + getval("tHX") + getval("rd") + getval("ad") + at; d.dutyoff = d1 + 4.0e-6; d = update_dutycycle(d); abort_dutycycle(d,10.0); // Set Phase Tables settable(phH90,4,table1); settable(phXhx,4,table2); settable(phHhx,4,table3); settable(phRec,4,table4); setreceiver(phRec); // Begin Sequence txphase(phXhx); decphase(phH90); obspwrf(getval("aXhx")); decpwrf(getval("aH90")); obsunblank(); decunblank(); _unblank34(); delay(d1); sp1on(); delay(2.0e-6); sp1off(); delay(2.0e-6); // H to X Cross Polarization decrgpulse(getval("pwH90"),phH90,0.0,0.0); decphase(phHhx); _cp_(hx,phHhx,phXhx); // Begin Acquisition _mpseqon(dec, phHhx); obsblank(); _blank34(); delay(getval("rd")); startacq(getval("ad")); acquire(np, 1/sw); endacq(); _mpseqoff(dec); obsunblank(); decunblank(); _unblank34(); }

void pulsesequence() { // Define Variables and Objects and Get Parameter Values DSEQ dec = getdseq("H"); strncpy(dec.t.ch,"dec",3); putCmd("chHtppm='dec'\n"); strncpy(dec.s.ch,"dec",3); putCmd("chHspinal='dec'\n"); //-------------------------------------- // Copy Current Parameters to Processed //------------------------------------- putCmd("groupcopy('current','processed','acquisition')"); // Dutycycle Protection DUTY d = init_dutycycle(); d.dutyon = getval("pwX90"); d.dutyoff = d1 + 4.0e-6; d.c1 = d.c1 + (!strcmp(dec.seq,"tppm")); d.c1 = d.c1 + ((!strcmp(dec.seq,"tppm")) && (dec.t.a > 0.0)); d.t1 = getval("rd") + getval("ad") + at; d.c2 = d.c2 + (!strcmp(dec.seq,"spinal")); d.c2 = d.c2 + ((!strcmp(dec.seq,"spinal")) && (dec.s.a > 0.0)); d.t2 = getval("rd") + getval("ad") + at; d = update_dutycycle(d); abort_dutycycle(d,10.0); // Set Phase Tables settable(phX90,4,table1); settable(phRec,4,table2); setreceiver(phRec); // Begin Sequence txphase(phX90); decphase(zero); obspwrf(getval("aX90")); obsunblank(); decunblank(); _unblank34(); delay(d1); sp1on(); delay(2.0e-6); sp1off(); delay(2.0e-6); // X Direct Polarization rgpulse(getval("pwX90"),phX90,0.0,0.0); // Begin Acquisition _dseqon(dec); obsblank(); _blank34(); delay(getval("rd")); startacq(getval("ad")); acquire(np, 1/sw); endacq(); _dseqoff(dec); obsunblank(); decunblank(); _unblank34(); }

pulsesequence() { settable(t1,4,phasecycle); dec3power(dpwr3); diplexer_override(0); delay(d1); dec3rgpulse(pw, t1, rof1, rof2); setreceiver(t1); }

pulsesequence() { // Define Variables and Objects and Get Parameter Values initval(getval("periods"),v2); //-------------------------------------- // Copy Current Parameters to Processed //------------------------------------- putCmd("groupcopy('current','processed','acquisition')"); // Set Phase Tables settable(phX90,4,table1); settable(phRec,4,table2); setreceiver(phRec); // Begin Sequence txphase(phX90); decphase(zero); obspwrf(getval("aX90")); obsunblank(); decunblank(); _unblank34(); delay(d1); xgate(1.0); sp1on(); delay(2.0e-6); sp1off(); delay(2.0e-6); // Apply a Rotorsync Delay rgpulse(getval("pwX90"),phX90,0.0,0.0); rotorsync(v2); rgpulse(getval("pwX90"),phX90,0.0,0.0); xgate(getval("xperiods")); rgpulse(getval("pwX90"),phX90,0.0,0.0); delay(10.0e-6); // X Direct Polarization rgpulse(getval("pwX90"),phX90,0.0,0.0); // Begin Acquisition obsblank(); _blank34(); delay(getval("rd")); startacq(getval("ad")); acquire(np, 1/sw); endacq(); obsunblank(); decunblank(); _unblank34(); }

void pulsesequence() { int rxgate; double pp, pplvl; pp = getval("pp"); pplvl = getval("pplvl"); rxgate = (rof1 == 0.0); if (rxgate) rof1 = 1.0e-6; /* phase switching time */ if (newdecamp) { if (rxgate) rof1 = 40.0e-6; } status(A); hsdelay(d1); status(B); settable(t1,4,phasecycle); pulse(pw, t1); delay(d2); if (newdecamp) { pplvl = getval("pplvl"); decpower(pplvl); /* sets DEC atten = pplvl */ } else { declvlon(); /* sets dhp = 255 level */ } simpulse(p1, pp, t1, t1, rof1, rof1); if (newdecamp) { decpower(dpwr); /* sets DEC atten = dpwr */ } else { declvloff(); } delay(d2); status(C); setreceiver(t1); }

void pulsesequence(){ //Define Variables and Get Parameter Values double pwTune = getval("pwTune"); pwTune = pwTune*6.0; at = pwTune*2.0; char atval[MAXSTR]; sprintf(atval,"at = %f\n", at); putCmd(atval); int chTune = (int) getval("chTune"); if ((chTune < 1) || (chTune > 4)) { abort_message("chTune(%d) must be between 1 and 4\n", chTune); } //-------------------------------------- // Copy Current Parameters to Processed //------------------------------------- putCmd("groupcopy('current','processed','acquisition')"); // Set Phase Tables settable(phTune,4,table1); settable(phRec,4,table2); setreceiver(t2); //Begin Sequence obspwrf(getval("aTune")); obsunblank(); decunblank(); _unblank34(); delay(d1); sp1on(); delay(2e-6); sp1off(); delay(2.0e-6); //Begin Phase Detected Pulse set4Tune(chTune,getval("gain")); delay(1.0e-4); ShapedXmtNAcquire("phtran",pwTune,phTune,6.0e-6,chTune); obsunblank(); decunblank(); _unblank34(); }

void pulsesequence() { /* equilibrium period */ status(A); hsdelay(d1); /* tau delay */ status(B); if (newdecamp) { decpower(tpwr); decrgpulse(p1, zero, rof1, rof2); decpower(dpwr); } else { declvlon(); decrgpulse(p1, zero, rof1, rof2); declvloff(); } hsdelay(d2); /* observe period */ status(C); settable(t1,4,phasecycle); if (newdecamp) { decpower(tpwr); decrgpulse(pw, t1, rof1, rof2); decpower(dpwr); } else { declvlon(); decrgpulse(pw, t1, rof1, rof2); declvloff(); } setreceiver(t1); }

void pulsesequence() { /* DECLARE VARIABLES */ char coshape[MAXSTR], /* dec pattern for CO decoupling */ fhfdwt1[MAXSTR], /* Flag to indicate half dwell start in t1 */ fhfdwt2[MAXSTR]; /* Flag to indicate half dwell start in t2 */ int satmove; double tauhc, /* 1 / 4*J[13C-H] [~1.5ms] */ tau1, /* t1-evolution variable */ tau2, /* t2-evolutionvariable */ pwca, /* PW90 for 13C nucleus */ pwco, /* PW90 for 13C carbonyl decoupling */ pwcalvl, /* power level for 13C pulses on dec1 */ pwcolvl, /* power level for C=O decoupling pulse */ jch, /* coupling for C-C (set to 40 Hz ) */ ncyc, /* # cycles through dipsi loop */ trim, /* trim pulse length(sec) */ dipsipwr, /* power level for 13C spin lock */ delta1,delta2; /* LOAD VARIABLES */ dipsipwr = getval("dipsipwr"); ncyc = getval("ncyc"); trim = getval("trim"); sw1 = getval("sw1"); sw2 = getval("sw2"); pwca = getval("pwca"); pwco = getval("pwco"); pwcalvl = getval("pwcalvl"); pwcolvl = getval("pwcolvl"); jch = getval("jch"); /* Use 152 [overestimate; true J ~140 ] */ getstr("coshape",coshape); getstr("fhfdwt1",fhfdwt1); getstr("fhfdwt2",fhfdwt2); /* calculate delays */ tauhc = (1.0/(4*jch)); delta1 = (1.0/(6.0*jch)); delta2 = (delta1); satmove = (fabs(tof - satfrq) >= 0.2); /* CHECK VALIDITY OF PARAMETER RANGE */ if((dm[A] != 'n') || (dm[B] != 'n')) { printf("dm should be 'nnn' or 'nny' "); psg_abort(1); } if((dm2[A] != 'n') || (dm2[B] != 'n') || (dm2[C] != 'n')) { printf("dm2 should be 'nnn' "); psg_abort(1); } if( satpwr > 20 ) { printf("SATPWR too large !!! "); psg_abort(1); } if( pwcolvl > 60 ) { printf("DCOPWR too large!"); psg_abort(1); } if( dpwr > 48 ) { printf("don't fry the probe, DCPWR too large! "); psg_abort(1); } if(( pwca > 2.5e-5 || pw > 2.5e-5 )) { printf("Pulsewidths [pw, pwca] must be shorter than 25 us. Abort."); psg_abort(1); } if ( pwco > 2.0e-3 ) { printf("PWCO must be less than 2 ms. Abort."); psg_abort(1); } if ( ncyc*217.33*p1 > 0.03) { printf("mixing time must be < 30ms! Abort."); psg_abort(1); } if (ncyc > 4) { printf("ncyc should be no greater than 4. Abort."); psg_abort(1); } /* INITIALIZE VARIABLES */ if(fhfdwt1[0] == 'y') tau1 = (d2 + 1/(2*sw1) ); else tau1 = d2; if(fhfdwt2[0] == 'y') tau2 = (d3 + 1/(2*sw2)); else tau2 = d3; initval(ncyc,v7); /* v7 is the dipsi loop counter */ settable(t21,1,psi1); settable(t11,2,phi1); settable(t12,1,phi2); settable(t13,1,phi3); settable(t14,1,phi4); settable(t15,8,phi5); settable(t16,8,phi6); settable(t10,8,rec); /* Phase table: phi1 = t11 = 0 1 phi2 = t12 = 0 phi3 = t13 = 0 0 0 0 0 0 0 0 phi3'= t14 = 1 1 1 1 1 1 1 1 phi5 = t15 = 0 0 1 1 2 2 3 3 phi6 = t16 = 0 0 0 0 2 2 2 2 THESE TO BE SOFTWARE-MODIFIED BASED ON t1, t2 VALUES: rec = t10 = 0 2 2 0 0 2 2 0 psi1 = t21 = 0 psi2 = t22 = 0 */ if(phase1==2) tsadd(t21,1,4); if(phase2==2) tsadd(t12,1,4); if(d2_index%2) { tsadd(t21,2,4); tsadd(t10,2,4); } if(d3_index%2) { tsadd(t12,2,4); tsadd(t10,2,4); } /* BEGIN ACTUAL PULSE SEQUENCE */ status(A); if (satmode[A] == 'y') { if(satmove) obsoffset(satfrq); obspower(satpwr); rgpulse(d1,zero,rof1,rof1); if(satmove) obsoffset(tof); } else { delay(d1); } rcvroff(); decphase(t13); obspower(tpwr); /* Set transmitter power for hard 1H pulses */ decpower(pwcalvl); status(B); rgpulse(pw,t21,1.0e-5,0.0); /* First 1H 90 degree pulse */ txphase(t11); if(tau1 < (2*pwca + pw)) /* first t1-value */ { delay(tauhc - 2*pwca - 1.0e-6); /* assuming pwca > pw */ decrgpulse(pwca,t13,0.0,0.0); /* 13C composite 180 deg pulse */ simpulse(2*pw, 2*pwca,t11, t14, 1.0e-6, 0.0); decrgpulse(pwca,t13,1.0e-6,0.0); /* 13C composite 180 deg pulse */ txphase(one); decphase(t12); delay(tauhc - 2*pwca - 1.0e-6); /* assuming pwca > pw */ } else { delay(tauhc + tau1/2 - 2*pwca -1.0e-6); /* t1-evol. plus pol. trans. */ decrgpulse(pwca,t13,0.0,0.0); /* 13C composite 180 deg pulse */ decrgpulse(2*pwca,t14,1.0e-6,0.0); /* 13C composite 180 deg pulse */ decrgpulse(pwca,t13,0.0,0.0); /* 13C composite 180 deg pulse */ delay(tau1/2 - 2*pwca - pw); /* continued t1-evol. */ rgpulse(2*pw,t11,0.0,0.0); /* proton echo pulse */ txphase(one); decphase(t12); delay(tauhc - pw); } simpulse(pw,pwca,one,t12,1.0e-6,0.2e-6); decphase(zero); decpower(pwcolvl); if(tau2/2 > POWER_DELAY) delay(tau2/2 - POWER_DELAY); /* t2 evolution */ else delay(tau2/2); decshaped_pulse(coshape,pwco,zero,0.0,0.0); decpower(pwcalvl); delay(delta1 - pwco - POWER_DELAY - WFG_START_DELAY - WFG_STOP_DELAY - pw); rgpulse(2*pw,zero,0.0,0.0); if (tau2/2 > (pw + pwca)) delay(tau2/2 - pw -pwca); /* t2 evolution */ else delay(tau2/2); decrgpulse(2*pwca,t15,0.0,0.0); decphase(t16); decpower(dipsipwr); delay(delta1 - pwca - POWER_DELAY); if (ncyc>0.0) { decrgpulse(trim,t16,0.0,0.0); starthardloop(v7); dipsi3a(); dipsi3b(); dipsi3b(); dipsi3a(); endhardloop(); } txphase(zero); decphase(zero); decpower(pwcalvl); delay(delta2 - POWER_DELAY - pwca); simpulse(2*pw,2*pwca,zero,zero,0.0,0.0); txphase(t13); delay(delta2); /* start reversed INEPT */ simpulse(pw,pwca,t13,zero,0.0,0.0); decphase(zero); txphase(zero); delay(tauhc - 2*pwca - 1.0e-6); /* delay = 1/4J */ decrgpulse(pwca,zero,0.0,0.0); simpulse(2*pw,2*pwca,zero,one,1.0e-6,0.0); decrgpulse(pwca,zero,1.0e-6,rof2); delay(tauhc - 2*pwca - 1.0e-6 - rof2); /* delay = 1/4J */ decrgpulse(pwca,zero,0.0,0.0); /* Filter out IySz terms */ decrgpulse(pwca,t16,1.0e-6,0.0); decpower(dpwr); /* Set power for decoupling */ /* BEGIN ACQUISITION */ setreceiver(t10); status(C); }

pulsesequence() { /* DECLARE VARIABLES */ char autocal[MAXSTR], /* auto-calibration flag */ fsat[MAXSTR], fscuba[MAXSTR], f1180[MAXSTR], /* Flag to start t1 @ halfdwell */ f2180[MAXSTR], /* Flag to start t2 @ halfdwell */ f3180[MAXSTR], /* Flag to start t3 @ halfdwell */ fco180[MAXSTR], /* Flag for checking sequence */ fca180[MAXSTR], /* Flag for checking sequence */ spca180[MAXSTR], /* string for the waveform Ca 180 */ spco180[MAXSTR], /* string for the waveform Co 180 */ spchirp[MAXSTR], /* string for the waveform reburp 180 */ ddseq[MAXSTR], /* 2H decoupling seqfile */ shp_sl[MAXSTR], /* string for seduce shape */ sel_flg[MAXSTR]; int phase, phase2, phase3, ni2, ni3, icosel, t1_counter, /* used for states tppi in t1 */ t2_counter, /* used for states tppi in t2 */ t3_counter; /* used for states tppi in t3 */ double tau1, /* t1 delay */ tau2, /* t2 delay */ tau3, /* t2 delay */ taua, /* ~ 1/4JNH = 2.25 ms */ taub, /* ~ 1/4JNH = 2.25 ms */ zeta, /* time for C'-N to refocuss set to 0.5*24.0 ms */ bigTN, /* nitrogen T period */ pwc90, /* PW90 for c nucleus @ d_c90 */ pwc180on, /* PW180 at @ d_c180 */ pwchirp, /* PW180 for ca nucleus @ d_creb */ pwc180off, /* PW180 at d_c180 + pad */ tsatpwr, /* low level 1H trans.power for presat */ d_c90, /* power level for 13C pulses(pwc90 = sqrt(15)/4delta) delta is the separation between Ca and Co */ d_c180, /* power level for 180 13C pulses (pwc180on=sqrt(3)/2delta */ d_chirp, sw1, /* sweep width in f1 */ sw2, /* sweep width in f2 */ sw3, /* sweep width in f3 */ pw_sl, /* pw90 for H selective pulse on water ~ 2ms */ phase_sl, /* phase for pw_sl */ tpwrsl, /* power level for square pw_sl */ pwDlvl, /* Power for D decoupling */ pwD, /* pw90 at pwDlvl */ pwC, pwClvl, /* C-13 calibration */ compC, pwN, /* PW90 for 15N pulse */ pwNlvl, /* high dec2 pwr for 15N hard pulses */ gstab, /* delay to compensate for gradient gt5 */ gt1, gt2, gt3, gt4, gt5, gt6, gt7, gt8, gt9, gzlvl1, gzlvl2, gzlvl3, gzlvl4, gzlvl5, gzlvl6, gzlvl7, gzlvl8, gzlvl9; /* LOAD VARIABLES */ getstr("autocal",autocal); getstr("fsat",fsat); getstr("fco180",fco180); getstr("fca180",fca180); getstr("f1180",f1180); getstr("f2180",f2180); getstr("f3180",f3180); getstr("fscuba",fscuba); getstr("ddseq",ddseq); getstr("shp_sl",shp_sl); getstr("sel_flg",sel_flg); taua = getval("taua"); taub = getval("taub"); zeta = getval("zeta"); bigTN = getval("bigTN"); tpwr = getval("tpwr"); tsatpwr = getval("tsatpwr"); dpwr = getval("dpwr"); pwN = getval("pwN"); pwNlvl = getval("pwNlvl"); pwD = getval("pwD"); pwDlvl = getval("pwDlvl"); phase = (int) ( getval("phase") + 0.5); phase2 = (int) ( getval("phase2") + 0.5); phase3 = (int) ( getval("phase3") + 0.5); sw1 = getval("sw1"); sw2 = getval("sw2"); sw3 = getval("sw3"); ni2 = getval("ni2"); ni3 = getval("ni3"); pw_sl = getval("pw_sl"); phase_sl = getval("phase_sl"); tpwrsl = getval("tpwrsl"); gstab = getval("gstab"); gt1 = getval("gt1"); if (getval("gt2") > 0) gt2=getval("gt2"); else gt2=gt1*0.1; gt3 = getval("gt3"); gt4 = getval("gt4"); gt5 = getval("gt5"); gt6 = getval("gt6"); gt7 = getval("gt7"); gt8 = getval("gt8"); gt9 = getval("gt9"); gzlvl1 = getval("gzlvl1"); gzlvl2 = getval("gzlvl2"); gzlvl3 = getval("gzlvl3"); gzlvl4 = getval("gzlvl4"); gzlvl5 = getval("gzlvl5"); gzlvl6 = getval("gzlvl6"); gzlvl7 = getval("gzlvl7"); gzlvl8 = getval("gzlvl8"); gzlvl9 = getval("gzlvl9"); if(autocal[0]=='n') { getstr("spca180",spca180); getstr("spco180",spco180); getstr("spchirp",spchirp); pwc90 = getval("pwc90"); pwc180on = getval("pwc180on"); pwc180off = getval("pwc180off"); d_c90 = getval("d_c90"); d_c180 = getval("d_c180"); pwchirp = getval("pwchirp"); d_chirp = getval("d_chirp"); } else { strcpy(spca180,"Phard180ca"); strcpy(spco180,"Phard180co"); strcpy(spchirp,"Pchirp180"); if (FIRST_FID) { pwC = getval("pwC"); compC = getval("compC"); pwClvl = getval("pwClvl"); co90 = pbox("cal", CO90, CO180ps, dfrq, pwC*compC, pwClvl); co180 = pbox("cal", CO180, CO180ps, dfrq, pwC*compC, pwClvl); ca180 = pbox(spca180, CA180, CA180ps, dfrq, pwC*compC, pwClvl); co180a = pbox(spco180, CO180a, CA180ps, dfrq, pwC*compC, pwClvl); chirp = pbox(spchirp, CHIRP, CHIRPps, dfrq, pwC*compC, pwClvl); } pwc90 = co90.pw; d_c90 = co90.pwr; pwc180on = co180.pw; d_c180 = co180.pwr; pwc180off = ca180.pw; pwchirp = chirp.pw; d_chirp = chirp.pwr; } /* LOAD PHASE TABLE */ settable(t1,2,phi1); settable(t2,2,phi2); settable(t3,4,phi3); settable(t4,1,phi4); settable(t5,4,phi5); settable(t6,4,rec); /* CHECK VALIDITY OF PARAMETER RANGES */ if( bigTN - (ni3-1)*0.5/sw3 - WFG3_START_DELAY < 0.2e-6 ) { text_error(" ni3 is too big\n"); text_error(" please set ni3 smaller or equal to %d\n", (int) ((bigTN -WFG3_START_DELAY)*sw3*2.0) +1 ); psg_abort(1); } if((dm[A] == 'y' || dm[B] == 'y' || dm[C] == 'y' || dm[D] == 'y' )) { text_error("incorrect dec1 decoupler flags! "); psg_abort(1); } if((dm2[A] == 'y' || dm2[B] == 'y' || dm2[C] == 'y' || dm2[D] == 'y')) { text_error("incorrect dec2 decoupler flags! Should be 'nnnn' "); psg_abort(1); } if( tsatpwr > 6 ) { text_error("TSATPWR too large !!! "); psg_abort(1); } if( dpwr > 46 ) { text_error("don't fry the probe, DPWR too large! "); psg_abort(1); } if( dpwr2 > 46 ) { text_error("don't fry the probe, DPWR2 too large! "); psg_abort(1); } if( dpwr3 > 50 ) { text_error("don't fry the probe, dpwr3 too large! "); psg_abort(1); } if( d_c90 > 62 ) { text_error("don't fry the probe, DHPWR too large! "); psg_abort(1); } if( pw > 200.0e-6 ) { text_error("dont fry the probe, pw too high ! "); psg_abort(1); } if( pwN > 200.0e-6 ) { text_error("dont fry the probe, pwN too high ! "); psg_abort(1); } if( pwc90 > 200.0e-6 ) { text_error("dont fry the probe, pwc90 too high ! "); psg_abort(1); } if( pwc180off > 200.0e-6 ) { text_error("dont fry the probe, pwc180 too high ! "); psg_abort(1); } if( gt3 > 2.5e-3 ) { text_error("gt3 is too long\n"); psg_abort(1); } if( gt1 > 10.0e-3 || gt2 > 10.0e-3 || gt4 > 10.0e-3 || gt5 > 10.0e-3 || gt6 > 10.0e-3 || gt7 > 10.0e-3 || gt8 > 10.0e-3 || gt9 > 10.0e-3) { text_error("gt values are too long. Must be < 10.0e-3 or gt11=50us\n"); psg_abort(1); } if((fca180[A] == 'y') && (ni2 > 1)) { text_error("must set fca180='n' to allow Calfa evolution (ni2>1)\n"); psg_abort(1); } if((fco180[A] == 'y') && (ni > 1)) { text_error("must set fco180='n' to allow CO evolution (ni>1)\n"); psg_abort(1); } /* Phase incrementation for hypercomplex 2D data */ if (phase == 2) tsadd(t1,1,4); if (phase2 == 2) tsadd(t5,1,4); if (phase3 == 2) { tsadd(t4, 2, 4); icosel = 1; } else icosel = -1; /* Set up f1180 tau1 = t1 */ tau1 = d2; if((f1180[A] == 'y') && (ni > 1)) { if (pwc180off > 2.0*pwN) tau1 += (1.0/(2.0*sw1) - 4.0*pwc90/PI - pwc180off - WFG3_START_DELAY - WFG3_STOP_DELAY - 4.0e-6 - 2.0*POWER_DELAY - 4.0e-6); else tau1 += (1.0/(2.0*sw1) - 4.0*pwc90/PI - 2.0*pwN - WFG3_START_DELAY - WFG3_STOP_DELAY - 4.0e-6 - 2.0*POWER_DELAY - 4.0e-6); if(tau1 < 0.2e-6) { tau1 = 0.4e-6; text_error("tau1 could be negative"); } } else { if (pwc180off > 2.0*pwN) tau1 = tau1 - 4.0*pwc90/PI - pwc180off - WFG3_START_DELAY - WFG3_STOP_DELAY - 4.0e-6 - 2.0*POWER_DELAY - 4.0e-6; else tau1 = tau1 - 4.0*pwc90/PI - 2.0*pwN - WFG3_START_DELAY - WFG3_STOP_DELAY - 4.0e-6 - 2.0*POWER_DELAY - 4.0e-6; if(tau1 < 0.2e-6) tau1 = 0.4e-6; } tau1 = tau1/2.0; /* Set up f2180 tau2 = t2 */ tau2 = d3; if((f2180[A] == 'y') && (ni2 > 1)) { if (pwc180off > 2.0*pwN) tau2 += ( 1.0 / (2.0*sw2) - 4.0*pwc90/PI - 4.0e-6 - 2.0*POWER_DELAY - WFG3_START_DELAY - pwc180off - WFG3_STOP_DELAY - 4.0e-6); else tau2 += ( 1.0 / (2.0*sw2) - 4.0*pwc90/PI - 4.0e-6 - 2.0*POWER_DELAY - WFG3_START_DELAY - 2.0*pwN - WFG3_STOP_DELAY - 4.0e-6); if(tau2 < 0.2e-6) { tau2 = 0.4e-6; text_error("tau2 could be negative"); } } else { if (pwc180off > 2.0*pwN) tau2 = tau2 - 4.0*pwc90/PI - 4.0e-6 - 2.0*POWER_DELAY - WFG3_START_DELAY - pwc180off - WFG3_STOP_DELAY - 4.0e-6; else tau2 = tau2 - 4.0*pwc90/PI - 4.0e-6 - 2.0*POWER_DELAY - WFG3_START_DELAY - 2.0*pwN - WFG3_STOP_DELAY - 4.0e-6; if(tau2 < 0.2e-6) tau2 = 0.4e-6; } tau2 = tau2/2.0; /* Set up f3180 tau3 = t3 */ tau3 = d4; if ((f3180[A] == 'y') && (ni3 > 1)) { tau3 += ( 1.0 / (2.0*sw3) ); if(tau3 < 0.2e-6) tau3 = 0.4e-6; } tau3 = tau3/2.0; /* Calculate modifications to phases for States-TPPI acquisition */ if( ix == 1) d2_init = d2 ; t1_counter = (int) ( (d2-d2_init)*sw1 + 0.5 ); if(t1_counter % 2) { tsadd(t1,2,4); tsadd(t6,2,4); } if( ix == 1) d3_init = d3 ; t2_counter = (int) ( (d3-d3_init)*sw2 + 0.5 ); if(t2_counter % 2) { tsadd(t5,2,4); tsadd(t6,2,4); } if( ix == 1) d4_init = d4 ; t3_counter = (int) ( (d4-d4_init)*sw3 + 0.5 ); if(t3_counter % 2) { tsadd(t2,2,4); tsadd(t6,2,4); } /* BEGIN ACTUAL PULSE SEQUENCE */ status(A); obsoffset(tof); decoffset(dof); /* set Dec1 carrier at Co */ obspower(tsatpwr); /* Set transmitter power for 1H presaturation */ decpower(d_chirp); /* Set Dec1 power for hard 13C pulses */ dec2power(pwNlvl); /* Set Dec2 power for 15N hard pulses */ /* Presaturation Period */ if (fsat[0] == 'y') { delay(2.0e-5); rgpulse(d1,zero,2.0e-6,2.0e-6); /* presaturation */ obspower(tpwr); /* Set transmitter power for hard 1H pulses */ delay(2.0e-5); if (fscuba[0] == 'y') { delay(2.2e-2); rgpulse(pw,zero,2.0e-6,0.0); rgpulse(2*pw,one,2.0e-6,0.0); rgpulse(pw,zero,2.0e-6,0.0); delay(2.2e-2); } } else { delay(d1); } obspower(tpwr); /* Set transmitter power for hard 1H pulses */ txphase(zero); dec2phase(zero); delay(1.0e-5); /* Begin Pulses */ status(B); rcvroff(); lk_hold(); delay(20.0e-6); initval(1.0,v2); obsstepsize(phase_sl); xmtrphase(v2); /* shaped pulse */ obspower(tpwrsl); shaped_pulse(shp_sl,pw_sl,one,4.0e-6,0.0); xmtrphase(zero); obspower(tpwr); txphase(zero); delay(4.0e-6); /* shaped pulse */ rgpulse(pw,zero,0.0,0.0); /* 90 deg 1H pulse */ delay(0.2e-6); zgradpulse(gzlvl5,gt5); delay(2.0e-6); delay(taua - gt5 - 2.2e-6); /* taua <= 1/4JNH */ sim3pulse(2*pw,0.0e-6,2*pwN,zero,zero,zero,0.0,0.0); txphase(three); dec2phase(zero); decphase(zero); delay(0.2e-6); zgradpulse(gzlvl5,gt5); delay(200.0e-6); delay(taua - gt5 - 200.2e-6 - 2.0e-6); if (sel_flg[A] == 'n') { rgpulse(pw,three,2.0e-6,0.0); delay(0.2e-6); zgradpulse(gzlvl3,gt3); delay(200.0e-6); dec2rgpulse(pwN,zero,0.0,0.0); delay( zeta ); dec2rgpulse(2.0*pwN,zero,0.0,0.0); decshaped_pulse(spchirp,pwchirp,zero,0.0,0.0); delay(zeta -WFG_START_DELAY -pwchirp -WFG_STOP_DELAY -2.0e-6); dec2rgpulse(pwN,zero,2.0e-6,0.0); } else { rgpulse(pw,one,2.0e-6,0.0); initval(1.0,v3); dec2stepsize(45.0); dcplr2phase(v3); delay(0.2e-6); zgradpulse(gzlvl3,gt3); delay(200.0e-6); dec2rgpulse(pwN,zero,0.0,0.0); dcplr2phase(zero); delay(1.34e-3 - SAPS_DELAY - 2.0*pw); rgpulse(pw,one,0.0,0.0); rgpulse(2.0*pw,zero,0.0,0.0); rgpulse(pw,one,0.0,0.0); delay( zeta - 1.34e-3 - 2.0*pw); dec2rgpulse(2.0*pwN,zero,0.0,0.0); decshaped_pulse(spchirp,pwchirp,zero,0.0,0.0); delay(zeta -WFG_START_DELAY -pwchirp -WFG_STOP_DELAY -2.0e-6); dec2rgpulse(pwN,zero,2.0e-6,0.0); } dec2phase(zero); decphase(t1); decpower(d_c90); delay(0.2e-6); zgradpulse(gzlvl8,gt8); delay(200.0e-6); decrgpulse(pwc90,t1,2.0e-6,0.0); /* t1 period for Co evolution begins */ if (fco180[A]=='n') { decpower(d_c180); delay(tau1); sim3shaped_pulse("",spca180,"",0.0,pwc180off,2.0*pwN,zero,zero,zero,4.0e-6,0.0); decpower(d_c90); delay(tau1); } else /* for checking sequence */ { decpower(d_c180); decrgpulse(pwc180on,zero,4.0e-6,0.0); decpower(d_c90); } /* t1 period for Co evolution ends */ decrgpulse(pwc90,zero,4.0e-6,0.0); decoffset(dof-(174-56)*dfrq); /* change Dec1 carrier to Ca (55 ppm) */ delay(0.2e-6); zgradpulse(gzlvl4,gt4); delay(150.0e-6); /* Turn on D decoupling using the third decoupler */ dec3phase(one); dec3power(pwDlvl); dec3rgpulse(pwD,one,4.0e-6,0.0); dec3phase(zero); dec3power(dpwr3); dec3unblank(); setstatus(DEC3ch, TRUE, 'w', FALSE, dmf3); /* Turn on D decoupling */ decrgpulse(pwc90,t5,2.0e-6,0.0); /* t2 period for Ca evolution begins */ if (fca180[A]=='n') { decphase(zero); dec2phase(zero); decpower(d_c180); delay(tau2); sim3shaped_pulse("",spco180,"",0.0,pwc180off,2.0*pwN,zero,zero,zero,4.0e-6,0.0); decpower(d_c90); delay(tau2); } else /* for checking sequence */ { decpower(d_c180); decrgpulse(pwc180on,zero,4.0e-6,0.0); decpower(d_c90); } /* t2 period for Ca evolution ends */ decrgpulse(pwc90,zero,4.0e-6,0.0); /* Turn off D decoupling */ setstatus(DEC3ch, FALSE, 'c', FALSE, dmf3); dec3blank(); dec3phase(three); dec3power(pwDlvl); dec3rgpulse(pwD,three,4.0e-6,0.0); /* Turn off D decoupling */ decoffset(dof); /* set carrier back to Co */ decpower(d_chirp); delay(0.2e-6); zgradpulse(gzlvl9,gt9); delay(150.0e-6); /* t3 period begins */ dec2rgpulse(pwN,t2,2.0e-6,0.0); dec2phase(t3); delay(bigTN - tau3); dec2rgpulse(2.0*pwN,t3,0.0,0.0); decshaped_pulse(spchirp,pwchirp,zero,0.0,0.0); txphase(zero); dec2phase(t4); delay(0.2e-6); zgradpulse(gzlvl1,gt1); delay(500.0e-6); delay(bigTN - WFG_START_DELAY - pwchirp - WFG_STOP_DELAY -gt1 -500.2e-6 -2.0*GRADIENT_DELAY); delay(tau3); sim3pulse(pw,0.0e-6,pwN,zero,zero,t4,0.0,0.0); /* t3 period ends */ decpower(d_c90); decrgpulse(pwc90,zero,4.0e-6,0.0); decoffset(dof-(174-56)*dfrq); decrgpulse(pwc90,zero,20.0e-6,0.0); delay(0.2e-6); zgradpulse(gzlvl6,gt6); delay(2.0e-6); dec2phase(zero); delay(taub - POWER_DELAY - 4.0e-6 - pwc90 - 20.0e-6 - pwc90 - gt6 - 2.2e-6); sim3pulse(2*pw,0.0e-6,2*pwN,zero,zero,zero,0.0,0.0); decoffset(dof); delay(0.2e-6); zgradpulse(gzlvl6,gt6); delay(200.0e-6); txphase(one); dec2phase(one); delay(taub - gt6 - 200.2e-6); sim3pulse(pw,0.0e-6,pwN,one,zero,one,0.0,0.0); delay(0.2e-6); zgradpulse(gzlvl7,gt7); delay(2.0e-6); txphase(zero); dec2phase(zero); delay(taub - gt7 - 2.2e-6); sim3pulse(2*pw,0.0e-6,2*pwN,zero,zero,zero,0.0,0.0); delay(0.2e-6); zgradpulse(gzlvl7,gt7); delay(200.0e-6); delay(taub - gt7 - 200.2e-6); sim3pulse(pw,0.0e-6,pwN,zero,zero,zero,0.0,0.0); delay(gt2 +gstab -0.5*(pwN -pw) -2.0*pw/PI); rgpulse(2*pw,zero,0.0,0.0); delay(2.0e-6); zgradpulse(icosel*gzlvl2,gt2); decpower(dpwr); dec2power(dpwr2); delay(gstab -2.0e-6 -2.0*GRADIENT_DELAY -2.0*POWER_DELAY); lk_sample(); /* BEGIN ACQUISITION */ status(C); setreceiver(t6); }

void pulsesequence() { /* DECLARE AND LOAD VARIABLES */ char ch90shape[MAXSTR], ch180shape[MAXSTR], exp_mode[MAXSTR], /* flag to run 3D, or 2D time-shared 15N TROSY /13C HSQC-SE*/ decCACO[MAXSTR], caco180shape[MAXSTR], f1180[MAXSTR], /* Flag to start t1 @ halfdwell */ f2180[MAXSTR], f3180[MAXSTR], f4180[MAXSTR]; /* do TROSY on N15 and H1 */ int icosel, max_pcyc; /* used to get n and p type */ double tpwrs, ni2=getval("ni2"), ni3=getval("ni3"), tau1, tau1p,tau2,tau3,tau3p, /*evolution times in indirect dimensions */ tauNH=getval("tauNH"), /* 1/(4Jhn)*/ tauCH=getval("tauCH"), /* 1/(4Jch) */ tauCH1= getval("tauCH1"), /* tauCH/2.0+tauNH/2.0,*/ /* 1/(8Jch) +1/(8Jnh) */ tauCH2= getval("tauCH2"), swC = getval("swC"), /* spectral widths in 13C methyls */ pwClvl = getval("pwClvl"), /* coarse power for C13 pulse */ pwC = getval("pwC"), /* C13 90 degree pulse length at pwClvl */ swN = getval("swN"), /* spectral widths in 15N */ pwNlvl = getval("pwNlvl"), /* power for N15 pulses */ pwN = getval("pwN"), /* N15 90 degree pulse length at pwNlvl */ ch90pwr=getval("ch90pwr"), ch90pw=getval("ch90pw"), ch90corr=getval("ch90corr"), ch90dres=getval("ch90dres"), ch90dmf=getval("ch90dmf"), ch180pw=getval("ch180pw"), ch180pwr=getval("ch180pwr"), caco180pw=getval("caco180pw"), caco180pwr=getval("caco180pwr"), mix=getval("mix"), tpwrsf_d = getval("tpwrsf_d"), /* fine power adustment for first soft pulse(down)*/ tpwrsf_u = getval("tpwrsf_u"), /* fine power adustment for second soft pulse(up) */ pwHs = getval("pwHs"), /* H1 90 degree pulse length at tpwrs */ compH =getval("compH"), gstab = getval("gstab"), gt0 = getval("gt0"), gt1 = getval("gt1"), gt2 = getval("gt2"), gt3 = getval("gt3"), gt4 = getval("gt4"), gt5 = getval("gt5"), gt6 = getval("gt6"), gt7 = getval("gt7"), gt8 = getval("gt8"), gt9 = getval("gt9"), gt10 = getval("gt10"), gzlvl0 = getval("gzlvl0"), gzlvl1 = getval("gzlvl1"), gzlvl2 = getval("gzlvl2"), gzlvl3 = getval("gzlvl3"), gzlvl4 = getval("gzlvl4"), gzlvl5 = getval("gzlvl5"), gzlvl6 = getval("gzlvl6"), gzlvl7 = getval("gzlvl7"), gzlvl8 = getval("gzlvl8"), gzlvl9 = getval("gzlvl9"), gzlvl10 = getval("gzlvl10"), gzlvl11 = getval("gzlvl11"); getstr("f1180",f1180); getstr("f2180",f2180); getstr("ch180shape",ch180shape); getstr("ch90shape",ch90shape); getstr("decCACO",decCACO); getstr("caco180shape",caco180shape); getstr("exp_mode",exp_mode); tpwrs = tpwr - 20.0*log10(pwHs/(compH*pw*1.69)); /*needs 1.69 times more*/ tpwrs = (int) (tpwrs); /*power than a square pulse */ if (tpwrsf_d<4095.0) tpwrs=tpwrs+6.0; /* add 6dB to let tpwrsf_d control fine power ~2048*/ if( (exp_mode[A]!='2') && (exp_mode[A]!='3') && (exp_mode[A]!='4') ) {text_error("invalid exp_mode, Should be either 2D or 3D or 4D\n "); psg_abort(1); } /* LOAD PHASE TABLE */ settable(t1,1,phi1); settable(t2,4,phi2); settable(t12,4,phi2); {tsadd(t12,2,4);} settable(t3,1,phi3); settable(t4,2,phi4); settable(t5,2,phi5); settable(t6,4,phi6); settable(t7,8,phi7); settable(t8,8,phi8); /* changing sign */ if( (exp_mode[A]=='4') && (exp_mode[C]=='a') ) {tsadd(t7,2,4); tsadd(t5,2,4); } settable(t21,1,psi1); /*trosy and SE hsqc in reverse INPET */ settable(t22,1,psi2); settable(t23,1,psi2c); if(exp_mode[A]=='2') {settable(t31,2,rec);} if(exp_mode[A]=='3') {settable(t31,4,rec);} if(exp_mode[A]=='4') {settable(t31,8,rec);} if((dm2[A] == 'y') || (dm2[B] == 'y') || (dm2[C] == 'y') || (dm2[D] == 'y')) { text_error("incorrect dec2 decoupler flags! Should be 'nnnn' "); psg_abort(1); } /* special case for swapping t2 and t3 for test purposes */ if( (exp_mode[A]=='4') && (exp_mode[B]=='x') && (ni3=1) ) { text_error("Acquiring t3 axis in ni2 dimension (instead of t2), set nt to 8! "); tau3 = 0.5*(d3_index/swC+0.5/swC)-pw-rof1 -pwC*2.0/M_PI ; /* increment corresponds to 13C increment */ tau3p = 0.5*(d3_index/swN+0.5/swN) -pw-rof1 -pwC -pwN*2.0/M_PI -tau3; if(d3_index % 2) { tsadd(t7,2,4); tsadd(t8,2,4); tsadd(t31,2,4); } if (phase2 == 2) {tsadd(t7 ,1,4); tsadd(t8 ,1,4);} tau2=0.0; } else { if (phase2 == 2) {tsadd(t2 ,1,4); tsadd(t12,1,4);} if (phase3 == 2) {tsadd(t7 ,1,4); tsadd(t8 ,1,4);} if(d3_index % 2) { tsadd(t2,2,4); tsadd(t12,2,4); tsadd(t31,2,4); } tau3 = 0.5*(d4_index/swC+0.5/swC)-pw -rof1 -pwC*2.0/M_PI ; /* increment corresponds to 13C increment */ tau3p = 0.5*(d4_index/swN+0.5/swN) -pw -rof1 -pwC -pwN*2.0/M_PI -tau3; if(d4_index % 2) { tsadd(t7,2,4); tsadd(t8,2,4); tsadd(t31,2,4); } tau2 = d3; tau2 += 0.0*(-pw*4.0/M_PI-rof1*2.0); if((f2180[A] == 'y') && (ni2 > 0.0)) {tau2 += ( 1.0 / (2.0*sw2) ); } if(tau2 < 0.2e-6) {tau2 = 0.0;} tau2 = tau2/2.0; } /* simultaneous Ntrosy-ChsqcSE, last part */ /* Phase incrementation for hypercomplex 2D data, States-Haberkorn element */ if (phase1 == 1) {icosel = 1; } else { tsadd(t21,2,4); tsadd(t22,2,4); tsadd(t23,2,4); icosel = -1; } if(d2_index % 2) { tsadd(t4,2,4); tsadd(t5,2,4); tsadd(t31,2,4); } /* ECHO-ANTIECHO + STATES-TPPI t1, t1' in TROSY/HSQC last step */ tau1 = 1.0*d2_index/swC; /* increment corresponds to 13C increment */ tau1p = 1.0*d2_index*(1.0/swN-1.0/swC); /* BEGIN PULSE SEQUENCE */ status(A); obspower(tpwr); decpower(pwClvl); dec2power(pwNlvl); txphase(zero); decphase(zero); dec2phase(zero); delay(d1); /* Destroy 13C magnetization*/ decrgpulse(pwC*1.0, zero, 0.0, 0.0); zgradpulse(-gzlvl0, gt0); delay(gstab); /* NOESY */ if(exp_mode[A]!='2') { /* 3-4D */ if(exp_mode[A]=='4') { /* full 4D */ /* t3 evolution, the very first HSQC */ /* Hz -> HzXz INEPT */ rgpulse(pw,two,rof1,rof1); /* 1H pulse excitation */ zgradpulse(gzlvl7, gt0); delay(tauCH-gt0); decrgpulse(pwC*2.0, zero, 0.0, 0.0); delay(tauNH -tauCH -pwC*2.0 ); sim3pulse(2.0*pw, 0.0, 2.0*pwN, zero, zero, zero, 0.0, 0.0); delay(tauNH - gt0 -gstab); zgradpulse(gzlvl7, gt0); delay(gstab); rgpulse(pw, one, rof1, rof1); /* water defoc-refoc */ delay(gstab); zgradpulse(gzlvl8, gt8); delay(gstab); /* t3 time */ if((ni3==0)) { dec2rgpulse(pwN,t7,0.0,0.0); dec2rgpulse(pwN,two,0.0,0.0); decrgpulse(pwC, t8, 0.0, 0.0); decrgpulse(pwC, two, 0.0, 0.0); rgpulse(pw*2.0, zero, rof1, rof1); delay(pwN*2.0+pwC*2.0); } else { dec2rgpulse(pwN,t7,0.0,0.0); delay(tau3p); decrgpulse(pwC, t8, 0.0, 0.0); delay(tau3); rgpulse(pw*2.0, zero, rof1, rof1); delay(tau3); decrgpulse(pwC, two, 0.0, 0.0); delay(tau3p); dec2rgpulse(pwN,two,0.0,0.0); } /* water defoc-refoc */ delay(gstab); zgradpulse(gzlvl8, gt8); delay(gstab); /* back inept, water to +Z */ rgpulse(pw,one,rof1,rof1); zgradpulse(gzlvl9, gt0); delay(tauCH-gt0); decrgpulse(pwC*2.0, zero, 0.0, 0.0); delay(tauNH -tauCH -pwC*2.0 ); sim3pulse(2.0*pw, 0.0, 2.0*pwN, zero, zero, zero, 0.0, 0.0); delay(tauNH - gt0 -gstab); zgradpulse(gzlvl9, gt0); delay(gstab); rgpulse(pw, two, rof1, rof1); /* purge */ zgradpulse(gzlvl10, gt10); delay(2.0*gstab); } /*end of full 4D */ /************************* t2 evolution, 1H and NOE****************************** */ /* for the case of no flipbacks in NOE part of the experiment, shift first pulse in t2 time by 45 deg and let water bring itself back at the end of mixing time by radiation dumping */ if( (exp_mode[D]=='t') ) { initval(1.0, v10); obsstepsize(45.0); xmtrphase(v10); } else { xmtrphase(zero); obspower(tpwrs); obspwrf(tpwrsf_d); shaped_pulse("H2Osinc",pwHs,t12,rof1,rof1); obspower(tpwr); obspwrf(4095.0); } rgpulse(pw,t2,rof1,rof1); xmtrphase(zero); /* SAPS_DELAY */ delay(tau2); decrgpulse(2.0*pwC,zero,0.0,0.0); dec2rgpulse(2.0*pwN,zero,0.0,0.0); delay(tau2); rgpulse(pw*2.0,zero,rof1,rof1); delay(pwN*2.0+pwC*2.0 + SAPS_DELAY); rgpulse(pw,zero,rof1,rof1); if( (exp_mode[D]!='t') ) { obspower(tpwrs); obspwrf(tpwrsf_u); shaped_pulse("H2Osinc",pwHs,zero,rof1,rof1); obspower(tpwr); obspwrf(4095.0); } /* NOESY period */ delay(mix-gt2-4.0*gstab ); zgradpulse(gzlvl2, gt2); delay(4.0*gstab); } /* end 3-4 D acquisition */ /* N-TROSY/C-HSQCse */ /* Hz -> HzXz INEPT */ rgpulse(pw,two,rof1,rof1); /* 1H pulse excitation */ zgradpulse(gzlvl0, gt0); delay(tauCH-gt0); decrgpulse(pwC*2.0, zero, 0.0, 0.0); delay(tauNH -tauCH -pwC*2.0 ); sim3pulse(2.0*pw, 0.0, 2.0*pwN, zero, zero, zero, 0.0, 0.0); delay(tauNH - gt0 -gstab); zgradpulse(gzlvl0, gt0); delay(gstab); rgpulse(pw, one, rof1, rof1); /* on HzXz now */ /* water flipback*/ obspower(tpwrs); obspwrf(tpwrsf_u); shaped_pulse("H2Osinc",pwHs,two,rof1,rof1); obspower(tpwr); obspwrf(4095.0); /* purge */ zgradpulse(gzlvl3, gt3); dec2phase(t4); delay(gstab*2.0); /* t1 (C) and t1+t1'(N) evolution */ dec2rgpulse(pwN, t4, 0.0, 0.0); delay(gt4+gstab + gt4+gstab + pwC*3.0 +2.0*(pwHs +2.0*rof1)); if(decCACO[A]=='y'){ delay(2.0*caco180pw);} dec2rgpulse(2.0*pwN, zero, 0.0, 0.0); delay(tau1p); /* t1 */ decrgpulse(pwC,t5,0.0,0.0); delay(tau1*0.5); if(decCACO[A]=='y') { decpower(caco180pwr); decshaped_pulse(caco180shape,caco180pw,zero, 0.0, 0.0); decpower(pwClvl); } obspower(ch180pwr); /*180 on methyls*/ shaped_pulse(ch180shape,ch180pw,zero,rof1,rof1); obspower(tpwr); delay(tau1*0.5); zgradpulse(gzlvl4, gt4); /*coding */ delay(gstab + pwHs -ch180pw -2.0*GRADIENT_DELAY -2.0*POWER_DELAY -WFG_START_DELAY- WFG_STOP_DELAY); decrgpulse(2.0*pwC,zero,0.0,0.0); if(decCACO[A]=='y') { decpower(caco180pwr); decshaped_pulse(caco180shape,caco180pw,zero, 0.0, 0.0); decpower(pwClvl); } /* delay(ch180pw+2.0*rof1);*/ zgradpulse(gzlvl5, gt4); delay(gstab - rof1 -2.0*GRADIENT_DELAY -2.0*POWER_DELAY -WFG_START_DELAY- WFG_STOP_DELAY); /*Water flipback (flipdown actually ) */ obspower(tpwrs); obspwrf(tpwrsf_d); shaped_pulse("H2Osinc",pwHs,three,rof1,rof1); obspower(tpwr); obspwrf(4095.0); /* reverse double INEPT */ sim3pulse(pw, pwC, 0.0, t21, t23, zero, rof1, rof1); /* rgpulse(pw, t21, rof1, rof1); */ zgradpulse(gzlvl11, gt1); delay(gstab); delay(tauCH1 -gt1 -gstab -2.0*pwC + (-2.0/M_PI*pwC-0.5*(pwN-pwC) +pwN)); decrgpulse(2.0*pwC,zero,0.0,0.0); delay(tauNH -tauCH1 - 0.65*(pw + pwN)-rof1 -(pwC-pw) -(-2.0/M_PI*pwC-0.5*(pwN-pwC) +pwN) ); sim3pulse(2.0*pw, 0.0, 2.0*pwN, zero, zero, zero, 0.0, 0.0); zgradpulse(gzlvl11, gt1); delay(gstab); delay(tauCH1-gt1 -gstab-2.0*pwC); decrgpulse(2.0*pwC,zero,0.0,0.0); delay(tauNH -1.3*pwN -tauCH1); sim3pulse(pw, pwC, pwN, one, zero, zero, 0.0, 0.0); zgradpulse(gzlvl1, gt1); delay(gstab); delay(tauCH2-2.0*pwC-gt1-gstab); decrgpulse(2.0*pwC,zero,0.0,0.0); delay(tauNH -1.3*pwN-tauCH2); sim3pulse(2.0*pw, 0.0, 2.0*pwN, zero, zero, zero, 0.0, 0.0); zgradpulse(gzlvl1, gt1); delay(gstab); delay(tauNH-1.6*pwN -POWER_DELAY -ch90pw*ch90corr -gt1-gstab +pwN*0.5 -PRG_START_DELAY); /* delay(ch90pw*0.5); sim3pulse(0.0,0.0, pwN, one, zero, t22, 0.0, 0.0); delay(ch90pw*0.5);*/ /* sim3shaped_pulse(ch90shape,"hard","hard",ch90pw,0.0, pwN, zero,zero, t22,0.0,0.0);*/ /* ch90corr is a fraction of ch90pw to correct for a 1H phase rollcaused by shaped 90 on CH3 protons for a sinc pulse ch90corr=0.41 seems to be good. */ obspower(ch90pwr); txphase(one); obsunblank(); xmtron(); obsprgon(ch90shape,1.0/ch90dmf,ch90dres); /*PRG_START_DELAY */ delay(ch90pw*ch90corr-pwN*0.5); dec2rgpulse(pwN, t22, 0.0, 0.0); delay(ch90pw*(1.0-ch90corr)-pwN*0.5); obsprgoff(); xmtroff(); obsblank(); /*PRG_STOP_DELAY */ obspower(tpwr); /*POWER_DEALY */ delay( gstab +gt6 +2.0*GRADIENT_DELAY +2.0*POWER_DELAY -0.65*pw -POWER_DELAY +pwN*0.5 -ch90pw*(1.0-ch90corr) -PRG_STOP_DELAY); rgpulse(2.0*pw, zero, rof1, rof1); dec2power(dpwr2); decpower(dpwr); /* 2.0*POWER_DELAY */ zgradpulse(gzlvl6*icosel, gt6); /* 2.0*GRADIENT_DELAY */ delay(gstab); status(C); setreceiver(t31); }

pulsesequence() { char f1180[MAXSTR], f2180[MAXSTR], mag_flg[MAXSTR]; /* y for magic angle, n for z-gradient only */ int icosel, t1_counter, t2_counter; double ni2, ratio, /* used to adjust t1 semi-constant time increment */ tau1, tau2, taua, /* ~ 1/4JCH = 1.5 ms - 1.7 ms] */ taub, /* ~ 3.3 ms */ bigTC, /* ~ 8 ms */ bigTCO, /* ~ 6 ms */ bigTN, /* ~ 12 ms */ tauc, /* ~ 5.4 ms */ taud, /* ~ 2.3 ms */ gstab, /* ~0.2 ms, gradient recovery time */ pwClvl, /* High power level for carbon on channel 2 */ pwC, /* C13 90 degree pulse length at pwClvl */ compH, /* Compression factor for H1 on channel 1 */ compC, /* Compression factor for C13 on channel 2 */ pwNlvl, /* Power level for Nitrogen on channel 3 */ pwN, /* N15 90 degree pulse lenght at pwNlvl */ maxpwCN, bw, ofs, ppm, /* bandwidth, offset, ppm - temporary Pbox parameters */ pwCa90, /*90 "offC13" pulse at Ca(56ppm) xmtr at CO(174ppm) */ pwCa180, /*180 "offC17" pulse at Ca(56ppm) xmtr at CO(174ppm) */ pwCO90, /* 90 "offC6" pulse at CO(174ppm) xmtr at CO(174ppm)*/ pwCO180, /* 180 "offC8" pulse at CO(174ppm) xmtr at CO(174ppm)*/ pwCab180, /* 180 "offC27" pulse at Cab(46ppm) xmtr at CO(174ppm)*/ tpwrHd, /* Power level for proton decoupling on channel 1 */ pwHd, /* H1 90 degree pulse lenth at tpwrHd. */ waltzB1 = getval("waltzB1"), /* waltz16 field strength (in Hz) */ phi_CO, /* phase correction for Bloch-Siegert effect on CO */ phi_Ca, /* phase correction for Bloch-Siegert effect on Ca */ gt1, gt2, gt3, gt4, gt5, gt6, gt7, gt0, gzlvl1, gzlvl2, gzlvl3, gzlvl4, gzlvl5, gzlvl6, /* N15 selection gradient level in DAC units */ gzlvl7, gzlvl0, /* H1 gradient level in DAC units */ gzcal, /* gradient calibration (gcal) */ dfCa180, dfCab180, dfC90, dfCa90, dfCO180; /* LOAD VARIABLES */ getstr("f1180",f1180); getstr("f2180",f2180); getstr("mag_flg", mag_flg); gzcal = getval("gzcal"); ni2 = getval("ni2"); taua = getval("taua"); taub = getval("taub"); tauc = getval("tauc"); bigTC = getval("bigTC"); bigTCO = getval("bigTCO"); bigTN = getval("bigTN"); taud = getval("taud"); gstab = getval("gstab"); pwClvl = getval("pwClvl"); pwC = getval("pwC"); compH = getval("compH"); compC = getval("compC"); pwNlvl = getval("pwNlvl"); pwN = getval("pwN"); phi_CO = getval("phi_CO"); phi_Ca = getval("phi_Ca"); gt1 = getval("gt1"); gt2 = getval("gt2"); gt3 = getval("gt3"); gt4 = getval("gt4"); gt5 = getval("gt5"); gt6 = getval("gt6"); gt7 = getval("gt7"); gt0 = getval("gt0"); gzlvl1 = getval("gzlvl1"); gzlvl2 = getval("gzlvl2"); gzlvl3 = getval("gzlvl3"); gzlvl4 = getval("gzlvl4"); gzlvl5 = getval("gzlvl5"); gzlvl6 = getval("gzlvl6"); gzlvl7 = getval("gzlvl7"); gzlvl0 = getval("gzlvl0"); setautocal(); /* activate auto-calibration flags */ if (autocal[0] == 'n') { pwCa90 = getval("pwCa90"); pwCa180 = getval("pwCa180"); pwCab180 = getval("pwCab180"); pwCO90 = getval("pwCO90"); pwCO180 = getval("pwCO180"); dfCa180 = (compC*4095.0*pwC*2.0*1.69)/pwCa180; /*power for "offC17" pulse*/ dfCab180 = (compC*4095.0*pwC*2.0*1.69)/pwCab180; /*power for "offC27" pulse*/ dfC90 = (compC*4095.0*pwC*1.69)/pwCO90; /*power for "offC6" pulse */ dfCa90 = (compC*4095.0*pwC)/pwCa90; /*power for "offC13" pulse*/ dfCO180 = (compC*4095.0*pwC*2.0*1.65)/pwCO180; /*power for "offC8" pulse */ dfCa90 = (int) (dfCa90 + 0.5); dfCa180 = (int) (dfCa180 + 0.5); dfC90 = (int) (dfC90 + 0.5); dfCO180 = (int) (dfCO180 + 0.5); dfCab180 = (int) (dfCab180 +0.5); /* power level and pulse time for WALTZ 1H decoupling */ pwHd = 1/(4.0 * waltzB1) ; tpwrHd = tpwr - 20.0*log10(pwHd/(compH*pw)); tpwrHd = (int) (tpwrHd + 0.5); } else { if(FIRST_FID) /* call Pbox */ { ppm = getval("dfrq"); bw = 118.0*ppm; ofs = -118.0*ppm; offC6 = pbox_make("offC6", "sinc90n", bw, 0.0, compC*pwC, pwClvl); offC8 = pbox_make("offC8", "sinc180n", bw, 0.0, compC*pwC, pwClvl); offC17 = pbox_make("offC17", "sinc180n", bw, ofs, compC*pwC, pwClvl); bw = 128.0*ppm; offC13 = pbox_make("offC13", "square90n", bw, ofs, compC*pwC, pwClvl); ofs = -128.0*ppm; offC27 = pbox_make("offC27", "sinc180n", bw, ofs, compC*pwC, pwClvl); bw = 2.8*7500.0; wz16 = pbox_Dcal("WALTZ16", 2.8*waltzB1, 0.0, compH*pw, tpwr); ofs_check(H1ofs, C13ofs, N15ofs, H2ofs); } dfC90 = offC6.pwrf; pwCO90 = offC6.pw; dfCO180 = offC8.pwrf; pwCO180 = offC8.pw; dfCa90 = offC13.pwrf; pwCa90 = offC13.pw; dfCa180 = offC17.pwrf; pwCa180 = offC17.pw; dfCab180 = offC27.pwrf; pwCab180 = offC27.pw; tpwrHd = wz16.pwr; pwHd = 1.0/wz16.dmf; } maxpwCN = 2.0*pwN; if (pwCab180 > pwN) maxpwCN = pwCab180; /* LOAD PHASE TABLE */ settable(t1,4,phi1); settable(t2,2,phi2); settable(t3,8,phi3); settable(t4,16,phi4); settable(t5,1,phi5); settable(t16,8,rec); /* CHECK VALIDITY OF PARAMETER RANGES */ if(ni > 64) { printf("ni is out of range. Should be: 14 to 64 ! \n"); psg_abort(1); } /* if(ni/sw1 > 2.0*(bigTCO)) { printf("ni is too big, should be < %f\n", sw1*2.0*(bigTCO)); psg_abort(1); } */ if(ni2/sw2 > 2.0*(bigTN - pwCO180)) { printf("ni2 is too big, should be < %f\n",2.0*sw2*(bigTN-pwCO180)); psg_abort(1); } if((dm[A] == 'y' || dm[B] == 'y' )) { printf("incorrect dec1 decoupler flags! Should be 'nnn' "); psg_abort(1); } if((dm2[A] == 'y' || dm2[B] == 'y')) { printf("incorrect dec2 decoupler flags! Should be 'nny' "); psg_abort(1); } if( dpwr > 50 ) { printf("don't fry the probe, DPWR too large! "); psg_abort(1); } /* Phase incrementation for hypercomplex 2D data */ if (phase1 == 1) { tsadd(t1, 1, 4); } if (phase2 == 2) { tsadd(t5,2,4); icosel = 1; } else icosel = -1; /* Set up f1180 tau1 = t1 */ tau1 = d2; if ((f1180[A] == 'y') && (ni > 1)) { tau1 += (1.0/(2.0*sw1)); } if(tau1 < 0.2e-6) tau1 = 0.0; tau1 = tau1/4.0; ratio = 2.0*bigTCO*sw1/((double) ni); ratio = (double)((int)(ratio*100.0))/100.0; if (ratio > 1.0) ratio = 1.0; if((dps_flag) && (ni > 1)) printf("ratio = %f => %f\n",2.0*bigTCO*sw1/((double) ni), ratio); /* Set up f2180 tau2 = t2 */ tau2 = d3; if ((f2180[A] == 'y') && (ni2 > 1)) { tau2 += (1.0/(2.0*sw2)); } if(tau2 < 0.2e-6) tau2 = 0.0; tau2 = tau2/4.0; /* Calculate modifications to phases for States-TPPI acquisition */ if( ix == 1) d2_init = d2 ; t1_counter = (int)((d2-d2_init)*sw1 + 0.5); if((t1_counter % 2)) { tsadd(t1,2,4); tsadd(t16,2,4); } if( ix == 1) d3_init = d3 ; t2_counter = (int)((d3-d3_init)*sw2 + 0.5); if((t2_counter % 2)) { tsadd(t2,2,4); tsadd(t16,2,4); } decstepsize(1.0); initval(phi_CO, v1); initval(phi_Ca, v2); /* BEGIN ACTUAL PULSE SEQUENCE */ status(A); delay(d1-1.0e-3); obsoffset(tof); decoffset(dof); obspower(tpwr); decpower(pwClvl); decpwrf(4095.0); dec2power(pwNlvl); txphase(zero); decphase(zero); dec2phase(zero); rcvroff(); if(gt6 > 0.2e-6) { delay(10.0e-6); decrgpulse(pwC, zero, 1.0e-6, 1.0e-6); delay(0.2e-6); zgradpulse(gzlvl6, gt6); } decpwrf(dfCa180); delay(1.0e-3); rgpulse(pw,zero,1.0e-6,1.0e-6); delay(2.0e-6); zgradpulse(gzlvl0,gt0); delay(taua - gt0 - 2.0e-6 - WFG_START_DELAY); simshaped_pulse("","offC17",2.0*pw,pwCa180,zero,zero,1.0e-6,1.0e-6); /* c13 offset on CO, slp 180 on Ca */ delay(taua - gt0 - 500.0e-6 - WFG_STOP_DELAY); zgradpulse(gzlvl0,gt0); txphase(one); delay(500.0e-6); rgpulse(pw, one, 1.0e-6, 1.0e-6); decphase(zero); delay(2.0e-6); zgradpulse(gzlvl3,gt3); obspower(tpwrHd); decpwrf(dfCa90); delay(200.0e-6); /* c13 offset on CO, slp 90 on Ca */ decshaped_pulse("offC13", pwCa90, zero, 0.0, 0.0); delay(taub -PRG_START_DELAY); obsprgon("waltz16", pwHd, 180.0); xmtron(); decpwrf(dfC90); decphase(t1); delay(bigTC -taub -SAPS_DELAY -PWRF_DELAY); /* c13 offset on CO, on-res 90 on CO */ decshaped_pulse("offC6", pwCO90, t1, 0.0, 0.0); /* CO EVOLUTION BEGINS */ decpwrf(dfCO180); decphase(zero); delay(bigTCO/2.0 +maxpwCN/2.0 +WFG_STOP_DELAY -2.0*pwCO90/PI -ratio*tau1); /* c13 offset on CO, on-res 180 on CO */ decshaped_pulse("offC8", pwCO180, zero, 0.0, 0.0); decpwrf(dfCab180); delay(bigTCO/2.0 +(2.0 -ratio)*tau1 -PRG_STOP_DELAY); xmtroff(); obsprgoff(); /* c13 offset on CO, slp 180 at Cab */ sim3shaped_pulse("","offC27","",0.0,pwCab180,2.0*pwN,zero,zero,zero,0.0,0.0); obsprgon("waltz16", pwHd, 180.0); xmtron(); decpwrf(dfCO180); delay(bigTCO/2.0 +(2.0 -ratio)*tau1 -PRG_START_DELAY); /* c13 offset on CO, on-res 180 on CO */ decshaped_pulse("offC8", pwCO180, zero, 0.0, 0.0); decpwrf(dfC90); dcplrphase(v1); delay(bigTCO/2.0 +maxpwCN/2.0 +WFG_STOP_DELAY -2.0*pwCO90/PI -ratio*tau1 -SAPS_DELAY); /* CO EVOLUTION ENDS */ decshaped_pulse("offC6", pwCO90, zero, 0.0, 0.0); /* c13 offset on CO, on-res 90 on CO */ decpwrf(dfCa90); decphase(t3); dcplrphase(v2); delay(bigTC -2.0*SAPS_DELAY -PWRF_DELAY); /* c13 offset on CO, slp 90 at Ca */ decshaped_pulse("offC13", pwCa90, t3, 0.0, 0.0); xmtroff(); decpwrf(dfCO180); decphase(zero); dcplrphase(zero); dec2phase(t2); delay(2.0e-5); zgradpulse(gzlvl4,gt4); delay(2.0e-6); obsprgon("waltz16", pwHd, 180.0); xmtron(); txphase(zero); delay(150.0e-6); dec2rgpulse(pwN, t2, 0.0, 0.0); /* N15 EVOLUTION BEGINS HERE */ delay(bigTN/2.0 -tau2); decshaped_pulse("offC8", pwCO180, zero, 0.0, 0.0); /* c13 offset on CO, on-res 180 on CO */ decpwrf(dfCa180); dec2phase(t4); delay(bigTN/2.0 -tau2); dec2rgpulse(2.0*pwN, t4, 0.0, 0.0); decshaped_pulse("offC17", pwCa180, zero, 0.0, 0.0); /* c13 offset on CO, slp 180 at Ca */ decpwrf(dfCO180); delay(bigTN/2.0 +tau2 -pwCa180 -WFG_START_DELAY -WFG_STOP_DELAY); decshaped_pulse("offC8", pwCO180, zero, 0.0, 0.0); /* c13 offset on CO, on-res 180 on CO */ delay(bigTN/2.0 +tau2 -tauc -PRG_STOP_DELAY); dec2phase(t5); xmtroff(); obsprgoff(); obspower(tpwr); if (mag_flg[A] == 'y') magradpulse(gzcal*gzlvl1, gt1); else zgradpulse(gzlvl1, gt1); delay(tauc -gt1 -2.0*GRADIENT_DELAY); /* N15 EVOLUTION ENDS HERE */ sim3pulse(pw,0.0, pwN, zero,zero, t5, 0.0, 0.0); dec2phase(zero); delay(2.0e-6); zgradpulse(0.8*gzlvl5, gt5); delay(taud - gt5 - 2.0e-6); sim3pulse(2.0*pw,0.0, 2.0*pwN, zero,zero, zero, 0.0, 0.0); delay(taud - gt5 - 500.0e-6); zgradpulse(0.8*gzlvl5, gt5); txphase(one); decphase(one); delay(500.0e-6); sim3pulse(pw,0.0, pwN, one,zero, one, 0.0, 0.0); delay(2.0e-6); txphase(zero); decphase(zero); zgradpulse(gzlvl5, gt5); delay(taud - gt5 - 2.0e-6); sim3pulse(2.0*pw,0.0, 2.0*pwN, zero,zero, zero, 0.0, 0.0); delay(taud - gt5 - 2.0*POWER_DELAY - 500.0e-6); zgradpulse(gzlvl5, gt5); decpower(dpwr); dec2power(dpwr2); delay(500.0e-6); rgpulse(pw, zero, 0.0, 0.0); delay(gstab +gt2 +2.0*GRADIENT_DELAY); rgpulse(2.0*pw, zero, 0.0, 0.0); if (mag_flg[A] == 'y') magradpulse(icosel*gzcal*gzlvl2, gt2); else zgradpulse(icosel*gzlvl2, gt2); delay(0.5*gstab); rcvron(); statusdelay(C, 0.5*gstab); setreceiver(t16); }

pulsesequence() { /* declare new variables */ double dutycycle, timeoff, cntct, pcrho, pdpd2, p180, pwx, srate, crossp, dipolr, tpwrm, rof2init, d2init, qpshft, lvlshft, ltoss, lpdp, lrof2; char pdp[MAXSTR], toss[MAXSTR], xpol[MAXSTR]; /* set variables */ cntct = getval("cntct"); p180 = getval("p180"); pcrho = getval("pcrho"); at = getval("at"); crossp = getval("crossp"); dipolr = getval("dipolr"); tpwrm = getval("tpwrm"); pwx = getval("pwx"); srate = getval("srate"); rof2init = getval("rof2"); d2init = getval("d2"); pdpd2 = getval("pdpd2"); getstr("xpol", xpol); getstr("pdp", pdp); getstr("toss", toss); /*adjust for propagation delays in the sequence*/ qpshft = 1.0e-6; lvlshft = 2.4e-6; ltoss = 0.0; lpdp = 0.0; lrof2 = 0.0; if (pdp[0]=='y') { lpdp = lpdp + lvlshft; } else { if (toss[0]=='y') { ltoss = ltoss + lvlshft; } else { lrof2 = lrof2 + lvlshft; } } /*adjust the rof2 delay and the d2 delay*/ rof2 = rof2init - lrof2; if (rof2 < 0.0) rof2 = 0.0; if (p180 > 0.0) { d2 = d2init - rof1; if (d2 < 0.0) d2 = 0.0; } /*set spin rate abort conditions for the sequence*/ if (toss[0]=='y') { if (((0.0773 / srate) - 2.0 * pwx - qpshft) < 0.0) { fprintf(stdout, "spin rate is too fast for TOSS!\n"); psg_abort(1); } } if (pdp[0]=='y') { if (((1.0 / srate) - pwx - pdpd2 - qpshft - lpdp) < 0.0) { fprintf(stdout, "pdpd2 is too long for spin rate!\n"); psg_abort(1); } } if (((toss[0] == 'y') || (pdp[0] == 'y')) && (srate < 500.0)) { fprintf(stdout, "spin rate is too low for toss or dipolar dephasing!\n"); psg_abort(1); } /*set abort conditions for high dutycycle and dm='y'*/ dutycycle = cntct + pcrho + pw + d1 + at; timeoff = d1; if (dm[2] != 'y') timeoff = timeoff + at; if (p180 > 0.0) { dutycycle = dutycycle + d2 + p180; timeoff = timeoff + d2; } if (toss[0] == 'y') { dutycycle = dutycycle + 2.142/srate; if (dm[2] != 'y') timeoff = timeoff + 2.142/srate - 8*pwx; } if (pdp[0] == 'y') { dutycycle = dutycycle + 2.0/srate; timeoff = timeoff + pdpd2; } dutycycle = timeoff/dutycycle; if ((dutycycle < 0.8) || (dm[0] == 'y')) { fprintf(stdout, "Duty cycle is %5.2f%%.\n", (1.0 - dutycycle) * 100.0); fprintf(stdout, "ABORT! The duty cycle must be less than 20%%.\n"); psg_abort(1); } /*begin pulse sequence*/ if (xpol[0] == 'n') { settable(t1,4,table1); settable(t3,4,table3); settable(t4,4,table4); status(A); setreceiver(t1); obs_pw_ovr(TRUE); dec_pw_ovr(TRUE); declvloff(); decpwrf(dipolr); obspwrf(tpwrm); delay(d1); if(p180 > 0.0) { decoff(); if (dm[1] == 'y') decon(); rgpulse(p180, zero, rof1, 0.0); delay(d2); } rcvroff(); rgpulse(pw, t1,rof1, 0.0); decoff(); if (dm[2] == 'y') decon(); } else { settable(t1,4,table1); settable(t2,4,table2); settable(t3,4,table3); settable(t4,4,table4); status(A); setreceiver(t1); obs_pw_ovr(TRUE); dec_pw_ovr(TRUE); declvloff(); decpwrf(crossp); obspwrf(tpwrm); txphase(t3); delay(d1); if(p180 > 0.0) { decrgpulse(p180, zero, rof1, 0.0); delay(d2); } rcvroff(); decphase(t2); delay(rof1); decon(); delay(pw - 0.8e-6); decphase(zero); delay(0.8e-6); xmtron(); delay(cntct); xmtroff(); /*optional spin lock for 13C T1rho*/ if (pcrho > 0.0) { decoff(); if (dm[1] == 'y') decon(); xmtron(); delay(pcrho); xmtroff(); } decpwrf(dipolr); decoff(); if (dm[2] == 'y') { decon(); } } /*optional interrupted decoupling for protonated carbon dephasing*/ if (pdp[0] == 'y') { decoff(); delay(pdpd2); decon(); delay((1.0 / srate) - pwx - pdpd2 - qpshft - lpdp); txphase(t3); delay(qpshft); xmtron(); delay(2.0 * pwx); xmtroff(); delay((1.0 / srate) - pwx); } /*optional pi pulses for suppression of sidebands - TOSS*/ if (toss[0] == 'y') { delay((0.1226 / srate) - pwx - qpshft - ltoss); txphase(t3); delay(qpshft); xmtron(); delay(2.0*pwx); xmtroff(); delay((0.0773 / srate) - 2.0 * pwx - qpshft); txphase(t4); delay(qpshft); xmtron(); delay(2.0 * pwx); xmtroff(); delay((0.2236 / srate) - 2.0 * pwx - qpshft); txphase(t3); delay(qpshft); xmtron(); delay(2.0 * pwx); xmtroff(); delay((1.0433 / srate) - 2.0 * pwx - qpshft); txphase(t4); delay(qpshft); xmtron(); delay(2.0 * pwx); xmtroff(); delay((0.7744 / srate) - pwx); } /*begin acquisition*/ delay(rof2); rcvron(); delay(alfa+1.0/(2.0*fb)); acquire(np,1/sw); decoff(); declvlon(); }

void pulsesequence() { /* DECLARE VARIABLES */ char f1180[MAXSTR],f2180[MAXSTR],satmode[MAXSTR]; int icosel,t1_counter,t2_counter,first_FID; double /* DELAYS */ tau1, /* t1/2 */ tau2, /* t2/2 */ /* COUPLINGS */ jhn = getval("jhn"), tauhn, jnco = getval("jnco"), taunco, jcoca = getval("jcoca"), taucoca, jnca = getval("jnca"), taunca, jhaca = getval("jhaca"), tauhaca, jcaha = getval("jcaha"), taucaha, jcacb = getval("jcacb"), taucacb, /* PULSES */ pwN = getval("pwN"), /* PW90 for N-nuc */ pwC = getval("pwC"), /* PW90 for C-nuc */ pwHs = getval("pwHs"), /* pw for water selective pulse at twprsl */ /* POWER LEVELS */ satpwr = getval("satpwr"), /* low power level for presat */ tpwrsf_d = getval("tpwrsf_d"), /* fine power level "down" flipback pulse*/ tpwrsf_u = getval("tpwrsf_u"), /* fine power level "up" flipback pulse*/ tpwrs, /* power level for selective pulse for water */ tpwrd,pwHd, /* power/pulse width for decoupling */ pwClvl = getval("pwClvl"), /* power level for C hard pulses */ compH = getval("compH"), /* compression factor */ compC = getval("compC"), /* compression factor */ pwNlvl = getval("pwNlvl"), /* power level for N hard pulses */ rf90onco, pw90onco, /* power level/width for CO 90 pulses */ rf180onco, pw180onco, /* power level/width for CO 180 pulses */ rf180offca, pw180offca, /* power level/width for off-res Ca 180 pulses */ /* CONSTANTS */ lambda = getval("lambda"), /* J scaling factor */ kappa, /* semi constant-time factor */ ni2=getval("ni2"), waltzB1 = getval("waltzB1"), /* waltz16 field strength (in Hz) */ /* GRADIENT DELAYS AND LEVES */ gt0 = getval("gt0"), /* gradient time */ gt1 = getval("gt1"), /* gradient time */ gt3 = getval("gt3"), /* gradient time */ gt5 = getval("gt5"), gstab = getval("gstab"), gzlvl0 = getval("gzlvl0"), /* level of gradient */ gzlvl1 = getval("gzlvl1"), gzlvl2 = getval("gzlvl2"), gzlvl3 = getval("gzlvl3"), gzlvl5 = getval("gzlvl5"); /* power level and pulse time for WALTZ 1H decoupling */ pwHd = 1/(4.0 * waltzB1); tpwrd = tpwr - 20.0*log10(pwHd/(compH*pw)); tpwrd = (int) (tpwrd + 0.5); /* LOAD VARIABLES */ getstr("satmode",satmode); getstr("f1180",f1180); getstr("f2180",f2180); /* check validity of parameter range */ if ((dm[A] == 'y' || dm[B] == 'y' || dm[C] == 'y' )) { printf("incorrect Dec1 decoupler flags! "); psg_abort(1); } if ((dm2[A] == 'y' || dm2[B] == 'y' || dm2[C] == 'y' )) { printf("incorrect Dec2 decoupler flags! "); psg_abort(1); } if ( satpwr > 8 ) { printf("satpwr too large !!! "); psg_abort(1); } if ( dpwr > 50 ) { printf("don't fry the probe, dpwr too large! "); psg_abort(1); } if ( dpwr2 > 50 ) { printf("don't fry the probe, dpwr2 too large! "); psg_abort(1); } /* LOAD VARIABLES */ settable(t1, 1, phi1); settable(t2, 1, phi2); settable(t3, 1, phi3); settable(t4, 2, phi4); settable(t5, 4, phi5); settable(t6, 8, phi6); settable(t7, 4, phi7); /* INITIALIZE VARIABLES */ tauhn = ((jhn != 0.0) ? 1/(4*(jhn)) : 2.75e-3); taunco = ((jnco !=0.0) ? 1/(4*(jnco)) : 16.6e-3); taucoca = ((jcoca !=0.0) ? 1/(4*(jcoca)) : 4.5e-3); taunca = ((jnca !=0.0) ? 1/(4*(jnca)) : 12e-3); tauhaca = ((jhaca !=0.0) ? 1/(4*(jhaca)) : 12e-3); taucaha = ((jcaha !=0.0) ? 1/(4*(jcaha)) : 12e-3); taucacb = ((jcacb !=0.0) ? 1/(4*(jcacb)) : 12e-3); if((getval("arraydim") < 1.5) || (ix==1)) first_FID = 1; else first_FID = 0; /* 90 degree pulse on CO, null at Ca 118ppm away */ pw90onco = sqrt(15.0)/(4.0*118.0*dfrq); rf90onco = (4095.0*pwC*compC)/pw90onco; rf90onco = (int) (rf90onco + 0.5); if(rf90onco > 4095.0) { if(first_FID) printf("insufficient power for pw90onco -> rf90onco (%.0f)\n", rf90onco); rf90onco = 4095.0; pw90onco = pwC; } /* 180 degree pulse on CO, null at Ca 118ppm away */ pw180onco = sqrt(3.0)/(2.0*118.0*dfrq); rf180onco = (4095.0*pwC*compC*2.0)/pw180onco; rf180onco = (int) (rf180onco + 0.5); if(rf180onco > 4095.0) { if(first_FID) printf("insufficient power for pw180onco -> rf180onco (%.0f)\n", rf180onco); rf180onco = 4095.0; pw180onco = pwC*2.0; } pw180offca = pw180onco; rf180offca = rf180onco; /* Phase incrementation for hypercomplex data */ if (phase1 == 2) /* Hypercomplex in t1 */ { tsadd(t4, 1, 4); } if (phase1 == 4) /* Hypercomplex in t1 */ { tsadd(t4, 1, 4); } kappa=(taunco - tauhn)/(0.5*ni2/sw2)-0.001; if (kappa > 1.0) { kappa=1.0-0.01; } if (phase2 == 1) /* Hypercomplex in t2 */ { icosel = -1; tsadd(t2, 2, 4); tsadd(t3, 2, 4); } else icosel = 1; if (ix == 1) printf("semi constant time factor %4.6f\n",kappa); /* calculate modification to phases based on current t1 values to achieve States-TPPI acquisition */ if (ix == 1) d2_init = d2; t1_counter = (int) ( (d2-d2_init)*sw1 + 0.5); if (t1_counter %2) /* STATES-TPPI */ { tsadd(t4,2,4); tsadd(t7,2,4); } if(ix==1) d3_init = d3; t2_counter = (int) ( (d3-d3_init)*sw2 + 0.5); if(t2_counter %2) { tsadd(t1,2,4); tsadd(t7,2,4); } /* set up so that get (-90,180) phase corrects in F1 if f1180 flag is y */ tau1 = d2; if (f1180[A] == 'y') tau1 += ( 1.0/(2.0*sw1)); tau1 = tau1/2.0; /* set up so that get (-90,180) phase corrects in F2 if f2180 flag is y */ tau2 = d3; if(f2180[A] == 'y') tau2 += ( 1.0/(2.0*sw2) ); tau2 = tau2/2.0; /* selective H20 one-lobe sinc pulse */ tpwrs = tpwr - 20.0*log10(pwHs/(compH*pw*1.69)); /*needs 1.69 times more*/ tpwrs = (int) (tpwrs); /*power than a square pulse */ if (tpwrsf_d<4095.0) tpwrs=tpwrs+6; /* nominal tpwrsf_d ~ 2048 */ /* tpwrsf_d,tpwrsf_u can be used to correct for radiation damping */ /* BEGIN ACTUAL PULSE SEQUENCE */ status(A); obspower(satpwr); /* Set power for presaturation */ decpower(pwClvl); /* Set decoupler1 power to pwClvl */ decpwrf(rf90onco); dec2power(pwNlvl); /* Set decoupler2 power to pwNlvl */ /* Presaturation Period */ if (satmode[0] == 'y') { rgpulse(d1,zero,rof1,0.0); obspower(tpwr); /* Set power for hard pulses */ } else { obspower(tpwr); /* Set power for hard pulses */ delay(d1); } status(B); rcvroff(); decpwrf(rf90onco); /* Set decoupler1 power to rf90onco */ sim3pulse(0.0,pw90onco,pwN,zero,zero,zero,rof1,rof1); /* 90 for 15N and 13C' */ zgradpulse(gzlvl0,gt0); delay(gstab); /* transfer from HN to N by INEPT */ /* shaped pulse for water flip-back */ obspower(tpwrs); obspwrf(tpwrsf_d); shaped_pulse("H2Osinc_d",pwHs,one,2.0e-6,0.0); obspower(tpwr); obspwrf(4095.0); /* shaped pulse */ rgpulse(pw,zero,rof1,0.0); zgradpulse(gzlvl0*1.3,gt0); delay(gstab); delay(tauhn - gt0 - gstab); /* 1/(4JHN) */ sim3pulse(2.0*pw,0.0,2.0*pwN,zero,zero,zero,rof1,rof1); delay(tauhn - gt0 - gstab); /* 1/(4JHN) */ zgradpulse(gzlvl0*1.3,gt0); delay(gstab); rgpulse(pw,three,rof1,0.0); /* 90 1H */ zgradpulse(gzlvl3,gt3); delay(gstab); decpwrf(rf180onco); /* Set decoupler power to rf180onco */ dec2rgpulse(pwN,zero,0.0,0.0); /* 90 15N */ /* start transfer from N to CO */ delay(5.5e-3 - pwHd - POWER_DELAY - PRG_START_DELAY); /* 1/(2JHN) */ obspower(tpwrd); rgpulse(pwHd,one,rof1,0.0); txphase(zero); obsprgon("waltz16", pwHd, 90.0); xmtron(); delay(taunco - 5.5e-3 - 0.5*pw180onco); /* 1/(4JNCO) - 1/(2JHN) */ sim3pulse(0.0,pw180onco,2.0*pwN,zero,zero,zero,rof1,rof1); delay(taunco - 5.5e-3 - 0.5*pw180onco); /* 1/(4JNCO) - 1/(2JHN) */ /* turn proton decoupling off */ xmtroff(); obsprgoff(); rgpulse(pwHd,three,rof1,0.0); obspower(tpwr); /* turned proton decoupling off */ delay(5.5e-3 - PRG_STOP_DELAY - pwHd - POWER_DELAY); /* 1/(2JHN) */ /* Start in-phase filter */ if (( phase1 == 1 || phase1 == 2)) { dec2rgpulse(pwN,one,0.0,0.0); /* 90 15N */ zgradpulse(gzlvl3*1.5,gt3); delay(gstab); /* shaped pulse WATER-FLIP-back */ obspower(tpwrs); obspwrf(tpwrsf_d); shaped_pulse("H2Osinc_d",pwHs,one,rof1,0.0); obspower(tpwr); obspwrf(4095.0); /* shaped pulse */ rgpulse(pw,one,rof1,0.0); /* 90 1H */ zgradpulse(gzlvl0*1.1,gt0); delay(gstab); delay(0.5*tauhn - gt0 - gstab); /* 1/(8JNH) */ dec2rgpulse(2.0*pwN,zero,0.0,0.0); /* 180 15N */ delay(0.5*tauhn - gt0 - gstab); /* 1/(8JNH) */ zgradpulse(gzlvl0*1.1,gt0); delay(gstab); rgpulse(2.0*pw,zero,rof1,rof1); /* 180 1H */ zgradpulse(gzlvl0*1.1,gt0); delay(gstab); delay(0.5*tauhn - gt0 -gstab); /* 1/(8JNH) */ dec2rgpulse(2.0*pwN,zero,0.0,0.0); /* 180 15N */ delay(0.5*tauhn - gt0 -gstab); /* 1/(8JNH) */ zgradpulse(gzlvl0*1.1,gt0); delay(gstab); rgpulse(pw,one,rof1,0.0); /* 90 1H */ /* shaped pulse WATER-FLIP-back */ obspower(tpwrs); obspwrf(tpwrsf_u); shaped_pulse("H2Osinc_u",pwHs,three,rof1,0.0); obspower(tpwr); obspwrf(4095.0); /* shaped pulse */ } /* start antiphase filter */ if (( phase1 == 3 || phase1 == 4 )) { dec2rgpulse(pwN,one,0.0,0.0); /* 90 15N */ zgradpulse(gzlvl3*1.5,gt3); delay(gstab); /* shaped pulse WATER-FLIP-back */ obspower(tpwrs); obspwrf(tpwrsf_d); shaped_pulse("H2Osinc_d",pwHs,zero,rof1,0.0); obspower(tpwr); obspwrf(4095.0); /* shaped pulse */ rgpulse(pw,zero,rof1,0.0); /* 90 1H */ zgradpulse(gzlvl0*0.9,gt0); delay(gstab); delay(tauhn - gt0 - gstab - 2.0*pwN); /* 1/(4JNH) */ sim3pulse(2.0*pw,0.0,2.0*pwN,zero,zero,zero,rof1,rof1); /* 180 1H and 15N */ delay(tauhn - gt0 - gstab - 2.0*pwN); /* 1/(4JNH) */ zgradpulse(gzlvl0*0.9,gt0); delay(gstab); rgpulse(pw,one,rof1,0.0); /* 90 1H */ /* shaped pulse WATER-FLIP-back */ obspower(tpwrs); obspwrf(tpwrsf_u); shaped_pulse("H2Osinc_u",pwHs,three,rof1,0.0); obspower(tpwr); obspwrf(4095.0); /* shaped pulse */ } /* End of filter section */ zgradpulse(gzlvl3*0.9,gt3); delay(gstab); rgpulse(2.0*pw,zero,rof1,0.0); decpwrf(rf90onco); /* Set decoupler power to rf90onco */ decrgpulse(pw90onco,t4,0.0,0.0); /* 90 for 13C */ /* record 13C' frequences and 13C'-15N coupling */ delay(tau1); decpwrf(rf180offca); /* Set decoupler power to rf180offca */ simshaped_pulse("","offC3",2.0*pw,pw180offca,zero,zero,rof1,rof1); /* 180 for 1H and 13CA */ decphase(zero); decpwrf(rf180onco); delay(tau1); if (lambda>0.0) { delay(lambda*tau1); sim3pulse(0.0,pw180onco,2.0*pwN,zero,t5,zero,0.0,0.0); delay(lambda*tau1); } else decrgpulse(pw180onco,t5,0.0,0.0); decpwrf(rf180offca); /* Set decoupler power to rf180offca */ simshaped_pulse("","offC3",0.0,pw180offca,zero,zero,0.0,0.0); /* 180 CA */ decphase(zero); decpwrf(rf90onco); /* start reverse transfer from CO to N by INEPT */ decrgpulse(pw90onco,zero,0.0,0.0); /* 90 for 13C' */ zgradpulse(gzlvl3*0.7,gt3); delay(gstab); dec2rgpulse(pwN,t1,0.0,0.0); /* 90 for 15N */ delay((taunco - tauhn) - kappa*tau2); /* 1/4J(NCO) - 1/4J(NH) - kt2/2 */ dec2rgpulse(2.0*pwN,zero,0.0,0.0); /* 180 for 15N */ delay((1-kappa)*tau2); /* (1-k)t2/2 */ decpwrf(rf180onco); decrgpulse(pw180onco,zero,0.0,0.0); /* 180 for 13C' */ decpwrf(rf180offca); delay(taunco - tauhn - gt1 - gstab - pw180onco - pw180offca - 3.0*POWER_DELAY); zgradpulse(gzlvl1,gt1); delay(gstab); decshaped_pulse("offC3",pw180offca,zero,0.0,0.0); delay(tau2); /* t2/2 */ /* start TROSY transfer from N to HN */ rgpulse(pw,t2,rof1,0.0); /* 180 for 1H */ zgradpulse(gzlvl5,gt5); delay(gstab); delay(tauhn - gt5 - gstab ); decpwrf(rf180onco); sim3pulse(2.0*pw,pw180onco,2.0*pwN,zero,zero,zero,rof1,rof1); delay(tauhn - gt5 - gstab ); zgradpulse(gzlvl5,gt5); delay(gstab); decpwrf(rf90onco); sim3pulse(pw,pw90onco,pwN,one,t6,zero,rof1,0.0); /* shaped pulse for water flip-back */ obspower(tpwrs); obspwrf(tpwrsf_u); shaped_pulse("H2Osinc_u",pwHs,t2,rof1,0.0); obspwrf(4095.0); obspower(tpwr); /* shaped pulse */ zgradpulse(gzlvl5*0.9,gt5); delay(gstab); delay(tauhn - gt5 - gstab - POWER_DELAY - pwHs); sim3pulse(2.0*pw,0.0,2.0*pwN,zero,zero,zero,rof1,rof1); delay(tauhn - gt5 - gstab ); decpower(dpwr); zgradpulse(gzlvl5*0.9,gt5); delay(gstab); dec2rgpulse(pwN,t3,0.0,0.0); /* 90 for 15N */ dec2power(dpwr2); delay((gt1/10.0) - pwN + gstab -POWER_DELAY); rgpulse(2.0*pw, zero, rof1, rof1); zgradpulse(gzlvl2*icosel,gt1/10.0); delay(gstab); /* acquire data */ status(C); setreceiver(t7); }

void pulsesequence() { /* DECLARE AND LOAD VARIABLES */ char f1180[MAXSTR], /* Flag to start t1 @ halfdwell */ mag_flg[MAXSTR], /*magic angle gradient*/ f2180[MAXSTR], /* Flag to start t2 @ halfdwell */ stCdec[MAXSTR], /* calls STUD+ waveforms from shapelib */ STUD[MAXSTR]; /* apply automatically calculated STUD decoupling */ int icosel1, /* used to get n and p type */ icosel2, t1_counter, /* used for states tppi in t1 */ t2_counter, /* used for states tppi in t2 */ ni2 = getval("ni2"); double tau1, /* t1 delay */ tau2, /* t2 delay */ del = getval("del"), /* time delays for CH coupling evolution */ BPdpwrspinlock, /* user-defined upper limit for spinlock(Hz) */ BPpwrlimits, /* =0 for no limit, =1 for limit */ del1 = getval("del1"), del2 = getval("del2"), /* STUD+ waveforms automatically calculated by macro "biocal" */ /* and string parameter stCdec calls them from your shapelib. */ stdmf, /* dmf for STUD decoupling */ studlvl, /* coarse power for STUD+ decoupling */ rf80 = getval("rf80"), /* rf in Hz for 80ppm STUD+ */ bw, ofs, ppm, /* temporary Pbox parameters */ pwClvl = getval("pwClvl"), /* coarse power for C13 pulse */ pwC = getval("pwC"), /* C13 90 degree pulse length at pwClvl */ rf0, /* maximum fine power when using pwC pulses */ /* p_d is used to calculate the isotropic mixing on the Cab region */ spinlock = getval("spinlock"), /* DIPSI-3 spinlock field strength in Hz */ p_d, /* 50 degree pulse for DIPSI-2 at rfd */ rfd, /* fine power for 7 kHz rf for 500MHz magnet */ ncyc = getval("ncyc"), /* no. of cycles of DIPSI-3 */ /* the following pulse lengths for SLP pulses are automatically calculated */ /* by the macro "ghcch_tocsy" . SLP pulse shapes, "offC10" etc are called */ /* directly from your shapelib. */ pwC10, /* 180 degree selective sinc pulse on CO(174ppm) */ pwZ, /* the largest of pwC10 and 2.0*pwN */ rf10, /* fine power for the pwC10 ("offC10") pulse */ compC = getval("compC"), /* adjustment for C13 amplifier compression */ pwNlvl = getval("pwNlvl"), /* power for N15 pulses */ pwN = getval("pwN"), /* N15 90 degree pulse length at pwNlvl */ sw1 = getval("sw1"), sw2 = getval("sw2"), gt1 = getval("gt1"), /* coherence pathway gradients */ gzcal = getval("gzcal"), /* G/cm to DAC coversion factor*/ gzlvl1 = getval("gzlvl1"), gzlvl2 = getval("gzlvl2"), gt3 = getval("gt3"), /* other gradients */ gt5 = getval("gt5"), gzlvl3 = getval("gzlvl3"), gzlvl4 = getval("gzlvl4"), gzlvl5 = getval("gzlvl5"), gzlvl6 = getval("gzlvl6"); getstr("STUD",STUD); getstr("mag_flg",mag_flg); getstr("f1180",f1180); getstr("f2180",f2180); strcpy(stCdec, "stCdec80"); stdmf = getval("dmf80"); studlvl = pwClvl + 20.0*log10(compC*pwC*4.0*rf80); studlvl = (int) (studlvl + 0.5); P_getreal(GLOBAL,"BPpwrlimits",&BPpwrlimits,1); P_getreal(GLOBAL,"BPdpwrspinlock",&BPdpwrspinlock,1); /* LOAD PHASE TABLE */ settable(t3,2,phi3); settable(t6,1,phi6); settable(t5,4,phi5); settable(t10,1,phi10); settable(t11,4,rec); /* INITIALIZE VARIABLES */ if (BPpwrlimits > 0.5) { if (spinlock > BPdpwrspinlock) { spinlock = BPdpwrspinlock; printf("spinlock too large, reset to user-defined limit (BPdpwrspinlock)"); psg_abort(1); } } if( dpwrf < 4095 ) { printf("reset dpwrf=4095 and recalibrate C13 90 degree pulse"); psg_abort(1); } /* maximum fine power for pwC pulses */ rf0 = 4095.0; setautocal(); /* activate auto-calibration flags */ if (autocal[0] == 'n') { /* "offC10": 180 degree one-lobe sinc pulse on CO, null at Ca 139ppm away */ pwC10 = getval("pwC10"); rf10 = (compC*4095.0*pwC*2.0*1.65)/pwC10; /* needs 1.65 times more */ rf10 = (int) (rf10 + 0.5); /* power than a square pulse */ if( pwC > (24.0e-6*600.0/sfrq) ) { printf("Increase pwClvl so that pwC < 24*600/sfrq"); psg_abort(1); } } else /* if autocal = 'y'(yes), 'q'(quiet), r(read), or 's'(semi) */ { if(FIRST_FID) /* call Pbox */ { ppm = getval("dfrq"); bw = 118.0*ppm; ofs = 139.0*ppm; offC10 = pbox_make("offC10", "sinc180n", bw, ofs, compC*pwC, pwClvl); if(dm3[B] == 'y') H2ofs = 3.2; ofs_check(H1ofs, C13ofs, N15ofs, H2ofs); } rf10 = offC10.pwrf; pwC10 = offC10.pw; } /* dipsi-3 decoupling on CbCa */ p_d = (5.0)/(9.0*4.0*spinlock); /* DIPSI-3 Field Strength */ rfd = (compC*4095.0*pwC*5.0)/(p_d*9.0); rfd = (int) (rfd + 0.5); ncyc = (int) (ncyc + 0.5); /* CHECK VALIDITY OF PARAMETER RANGES */ if( gt1 > 0.5*del - 1.0e-4) { printf(" gt1 is too big. Make gt1 less than %f.\n", (0.5*del - 1.0e-4)); psg_abort(1); } if( dm[A] == 'y' ) { printf("incorrect dec1 decoupler flag! Should be 'nny' or 'nnn' "); psg_abort(1); } if((dm2[A] == 'y' || dm2[C] == 'y')) { printf("incorrect dec2 decoupler flags! Should be 'nnn' "); psg_abort(1); } if((dm3[A] == 'y' || dm3[C] == 'y')) { printf("incorrect dec3 decoupler flags! Should be 'nnn' or 'nyn' "); psg_abort(1); } if( dpwr > 52 ) { printf("don't fry the probe, DPWR too large! "); psg_abort(1); } if( pw > 50.0e-6 ) { printf("dont fry the probe, pw too high ! "); psg_abort(1); } if( pwN > 100.0e-6 ) { printf("dont fry the probe, pwN too high ! "); psg_abort(1); } /* PHASES AND INCREMENTED TIMES */ /* Phase incrementation for hypercomplex 2D data, States-Haberkorn element */ icosel1 = -1; icosel2 = -1; if (phase1 == 2) { tsadd(t6,2,4); icosel1 = -1*icosel1; } if (phase2 == 2) { tsadd(t10,2,4); icosel2 = -1*icosel2; tsadd(t6,2,4); } /* Set up f1180 */ tau1 = d2; if((f1180[A] == 'y') && (ni > 1.0)) { tau1 += ( 1.0 / (2.0*sw1) ); if(tau1 < 0.2e-6) tau1 = 0.0; } tau1 = tau1/2.0; /* Set up f2180 */ tau2 = d3; if((f2180[A] == 'y') && (ni2 > 1.0)) { tau2 += ( 1.0 / (2.0*sw2) ); if(tau2 < 0.2e-6) tau2 = 0.0; } tau2 = tau2/2.0; /* Calculate modifications to phases for States-TPPI acquisition */ if( ix == 1) d2_init = d2; t1_counter = (int) ( (d2-d2_init)*sw1 + 0.5 ); if(t1_counter % 2) { tsadd(t3,2,4); tsadd(t11,2,4); } if( ix == 1) d3_init = d3; t2_counter = (int) ( (d3-d3_init)*sw2 + 0.5 ); if(t2_counter % 2) { tsadd(t5,2,4); tsadd(t11,2,4); } /* BEGIN PULSE SEQUENCE */ status(A); if ( dm3[B] == 'y' ) lk_sample(); if ((ni/sw1-d2)>0) delay(ni/sw1-d2); /*decreases as t1 increases for const.heating*/ if ((ni2/sw2-d3)>0) delay(ni2/sw2-d3); /*decreases as t2 increases for const.heating*/ delay(d1); if ( dm3[B] == 'y' ) { lk_hold(); lk_sampling_off();} /*freezes z0 correction, stops lock pulsing*/ rcvroff(); obspower(tpwr); decpower(pwClvl); dec2power(pwNlvl); decpwrf(rf0); obsoffset(tof); txphase(t3); delay(1.0e-5); decrgpulse(pwC, zero, 0.0, 0.0); /*destroy C13 magnetization*/ zgradpulse(gzlvl1, 0.5e-3); delay(1.0e-4); decrgpulse(pwC, one, 0.0, 0.0); zgradpulse(0.7*gzlvl1, 0.5e-3); delay(5.0e-4); if ( dm3[B] == 'y' ) /* begins optional 2H decoupling */ { dec3rgpulse(1/dmf3,one,10.0e-6,2.0e-6); dec3unblank(); dec3phase(zero); delay(2.0e-6); setstatus(DEC3ch, TRUE, 'w', FALSE, dmf3); } rgpulse(pw, t3, 0.0, 0.0); /* 1H pulse excitation */ decphase(zero); delay(0.5*del + tau1 - 2.0*pwC); decrgpulse(2.0*pwC, zero, 0.0, 0.0); txphase(zero); delay(tau1); rgpulse(2.0*pw, zero, 0.0, 0.0); if (mag_flg[A] == 'y') { magradpulse(icosel1*gzcal*gzlvl1,0.1*gt1); } else { zgradpulse(icosel1*gzlvl1, 0.1*gt1); } decphase(t5); delay(0.5*del - 0.1*gt1); simpulse(pw, pwC, zero, t5, 0.0, 0.0); zgradpulse(gzlvl3, gt3); decphase(zero); delay(0.5*del2 - gt3); simpulse(2.0*pw, 2.0*pwC, zero, zero, 0.0, 0.0); zgradpulse(gzlvl3, gt3); txphase(t6); decphase(one); delay(0.5*del2 - gt3); simpulse(pw, pwC, t6, one, 0.0, 0.0); zgradpulse(gzlvl4, gt3); txphase(zero); decphase(zero); delay(0.5*del1 - gt3); simpulse(2.0*pw, 2.0*pwC, zero, zero, 0.0, 0.0); zgradpulse(gzlvl4, gt3); delay(0.5*del1 - gt3); decrgpulse(pwC, zero, 0.0, 0.0); decpwrf(rfd); delay(2.0e-6); initval(ncyc, v2); starthardloop(v2); decrgpulse(4.9*p_d,zero,0.0,0.0); decrgpulse(7.9*p_d,two,0.0,0.0); decrgpulse(5.0*p_d,zero,0.0,0.0); decrgpulse(5.5*p_d,two,0.0,0.0); decrgpulse(0.6*p_d,zero,0.0,0.0); decrgpulse(4.6*p_d,two,0.0,0.0); decrgpulse(7.2*p_d,zero,0.0,0.0); decrgpulse(4.9*p_d,two,0.0,0.0); decrgpulse(7.4*p_d,zero,0.0,0.0); decrgpulse(6.8*p_d,two,0.0,0.0); decrgpulse(7.0*p_d,zero,0.0,0.0); decrgpulse(5.2*p_d,two,0.0,0.0); decrgpulse(5.4*p_d,zero,0.0,0.0); decrgpulse(0.6*p_d,two,0.0,0.0); decrgpulse(4.5*p_d,zero,0.0,0.0); decrgpulse(7.3*p_d,two,0.0,0.0); decrgpulse(5.1*p_d,zero,0.0,0.0); decrgpulse(7.9*p_d,two,0.0,0.0); decrgpulse(4.9*p_d,two,0.0,0.0); decrgpulse(7.9*p_d,zero,0.0,0.0); decrgpulse(5.0*p_d,two,0.0,0.0); decrgpulse(5.5*p_d,zero,0.0,0.0); decrgpulse(0.6*p_d,two,0.0,0.0); decrgpulse(4.6*p_d,zero,0.0,0.0); decrgpulse(7.2*p_d,two,0.0,0.0); decrgpulse(4.9*p_d,zero,0.0,0.0); decrgpulse(7.4*p_d,two,0.0,0.0); decrgpulse(6.8*p_d,zero,0.0,0.0); decrgpulse(7.0*p_d,two,0.0,0.0); decrgpulse(5.2*p_d,zero,0.0,0.0); decrgpulse(5.4*p_d,two,0.0,0.0); decrgpulse(0.6*p_d,zero,0.0,0.0); decrgpulse(4.5*p_d,two,0.0,0.0); decrgpulse(7.3*p_d,zero,0.0,0.0); decrgpulse(5.1*p_d,two,0.0,0.0); decrgpulse(7.9*p_d,zero,0.0,0.0); decrgpulse(4.9*p_d,two,0.0,0.0); decrgpulse(7.9*p_d,zero,0.0,0.0); decrgpulse(5.0*p_d,two,0.0,0.0); decrgpulse(5.5*p_d,zero,0.0,0.0); decrgpulse(0.6*p_d,two,0.0,0.0); decrgpulse(4.6*p_d,zero,0.0,0.0); decrgpulse(7.2*p_d,two,0.0,0.0); decrgpulse(4.9*p_d,zero,0.0,0.0); decrgpulse(7.4*p_d,two,0.0,0.0); decrgpulse(6.8*p_d,zero,0.0,0.0); decrgpulse(7.0*p_d,two,0.0,0.0); decrgpulse(5.2*p_d,zero,0.0,0.0); decrgpulse(5.4*p_d,two,0.0,0.0); decrgpulse(0.6*p_d,zero,0.0,0.0); decrgpulse(4.5*p_d,two,0.0,0.0); decrgpulse(7.3*p_d,zero,0.0,0.0); decrgpulse(5.1*p_d,two,0.0,0.0); decrgpulse(7.9*p_d,zero,0.0,0.0); decrgpulse(4.9*p_d,zero,0.0,0.0); decrgpulse(7.9*p_d,two,0.0,0.0); decrgpulse(5.0*p_d,zero,0.0,0.0); decrgpulse(5.5*p_d,two,0.0,0.0); decrgpulse(0.6*p_d,zero,0.0,0.0); decrgpulse(4.6*p_d,two,0.0,0.0); decrgpulse(7.2*p_d,zero,0.0,0.0); decrgpulse(4.9*p_d,two,0.0,0.0); decrgpulse(7.4*p_d,zero,0.0,0.0); decrgpulse(6.8*p_d,two,0.0,0.0); decrgpulse(7.0*p_d,zero,0.0,0.0); decrgpulse(5.2*p_d,two,0.0,0.0); decrgpulse(5.4*p_d,zero,0.0,0.0); decrgpulse(0.6*p_d,two,0.0,0.0); decrgpulse(4.5*p_d,zero,0.0,0.0); decrgpulse(7.3*p_d,two,0.0,0.0); decrgpulse(5.1*p_d,zero,0.0,0.0); decrgpulse(7.9*p_d,two,0.0,0.0); endhardloop(); dec2phase(zero); decphase(zero); txphase(zero); decpwrf(rf10); delay(tau2); /* WFG3_START_DELAY */ sim3shaped_pulse("", "offC10", "", 2.0*pw, pwC10, 2.0*pwN, zero, zero, zero, 0.0, 0.0); if(pwC10>2.0*pwN) pwZ=0.0; else pwZ=2.0*pwN - pwC10; delay(tau2); decpwrf(rf0); if (mag_flg[A] == 'y') { magradpulse(-icosel2*gzcal*gzlvl2, 1.8*gt1); } else { zgradpulse(-icosel2*gzlvl2, 1.8*gt1); } delay(2.02e-4); decrgpulse(2.0*pwC, zero, 0.0, 0.0); decpwrf(rf10); if (mag_flg[A] == 'y') { magradpulse(icosel2*gzcal*gzlvl2, 1.8*gt1); } else { zgradpulse(icosel2*gzlvl2, 1.8*gt1); } delay(2.0e-4 + WFG3_START_DELAY + pwZ); decshaped_pulse("offC10", pwC10, zero, 0.0, 0.0); decpwrf(rf0); decrgpulse(pwC, zero, 2.0e-6, 0.0); zgradpulse(gzlvl5, gt5); delay(0.5*del1 - gt5); simpulse(2.0*pw, 2.0*pwC, zero, zero, 0.0, 0.0); zgradpulse(gzlvl5, gt5); txphase(one); decphase(t10); delay(0.5*del1 - gt5); simpulse(pw, pwC, one, t10, 0.0, 0.0); zgradpulse(gzlvl6, gt5); txphase(zero); decphase(zero); delay(0.5*del2 - gt5); simpulse(2.0*pw, 2.0*pwC, zero, zero, 0.0, 0.0); zgradpulse(gzlvl6, gt5); delay(0.5*del2 - gt5); simpulse(pw, pwC, zero, zero, 0.0, 0.0); delay(0.5*del - 0.5*pwC); simpulse(2.0*pw,2.0*pwC, zero, zero, 0.0, 0.0); if (mag_flg[A] == 'y') magradpulse(gzcal*gzlvl1, gt1); else zgradpulse(gzlvl1, gt1); rcvron(); if ((STUD[A]=='n') && (dm[C] == 'y')) decpower(dpwr); if ( dm3[B] == 'y' ) /* turns off 2H decoupling */ { delay(0.5*del-40.0e-6 -gt1 -1/dmf3); setstatus(DEC3ch, FALSE, 'c', FALSE, dmf3); dec3rgpulse(1/dmf3,three,2.0e-6,2.0e-6); dec3blank(); lk_autotrig(); /* resumes lock pulsing */ lk_sample(); if (mag_flg[A] == 'y') statusdelay(C,40.0e-6 - 2.0*VAGRADIENT_DELAY - POWER_DELAY); else statusdelay(C,40.0e-6 - 2.0*GRADIENT_DELAY - POWER_DELAY); } else { delay(0.5*del-40.0e-6 -gt1); if (mag_flg[A] == 'y') statusdelay(C,40.0e-6 - 2.0*VAGRADIENT_DELAY - POWER_DELAY); else statusdelay(C,40.0e-6 - 2.0*GRADIENT_DELAY - POWER_DELAY); } if ((STUD[A]=='y') && (dm[C] == 'y')) {decpower(studlvl); decunblank(); decon(); decprgon(stCdec,1/stdmf, 1.0); startacq(alfa); acquire(np, 1.0/sw); decprgoff(); decoff(); decblank(); } setreceiver(t11); }

pulsesequence() { // Define Variables and Objects and Get Parameter Values double aXfam2 = getval("aXfam2"); double pw1Xfam2 = getval("pw1Xfam2"); double pw2Xfam2 = getval("pw2Xfam2"); double pw3Xfam2 = getval("pw3Xfam2"); double pw4Xfam2 = getval("pw4Xfam2"); double nXfam2 = getval("nXfam2"); initval(nXfam2,v4); putCmd("pw2Xmqmas=pwXfam1"); // Sequence uses pwXfam1 and sets pw2Xmqmas double d2init = getval("d2"); // Define the Split d2 in the Pulse Sequence double ival = getval("ival"); double d20 = 1.0; double d21 = 0.0; double d22 = 0.0; if (ival == 1.5) { d20 = 9.0*d2init/16.0; d21 = 7.0*d2init/16.0; d22 = 0.0; } else if (ival == 2.5) { d20 = 12.0*d2init/31.0; d21 = 0.0*d2init/31.0; d22 = 19.0*d2init/31.0; } else { d20 = 1.0*d2init; d21 = 0.0*d2init; d22 = 0.0*d2init; } double tXechselinit = getval("tXechsel"); // Adjust the selective echo delay for the double tXechsel = tXechselinit - 3.0e-6; // attenuator switch time. if (tXechsel < 0.0) tXechsel = 0.0; DSEQ dec = getdseq("H"); strncpy(dec.t.ch,"dec",3); putCmd("chHtppm='dec'\n"); strncpy(dec.s.ch,"dec",3); putCmd("chHspinal='dec'\n"); // Set Constant-time Period for d2. if (d2_index == 0) d2_init = getval("d2"); double d2_ = (ni - 1)/sw1 + d2_init; putCmd("d2acqret = %f\n",roundoff(d2_,12.5e-9)); putCmd("d2dwret = %f\n",roundoff(1.0/sw1,12.5e-9)); //-------------------------------------- // Copy Current Parameters to Processed //------------------------------------- putCmd("groupcopy('current','processed','acquisition')"); // Dutycycle Protection DUTY d = init_dutycycle(); d.dutyon = getval("pw1Xmqmas") + nXfam2*(pw1Xfam2 + pw2Xfam2 + pw3Xfam2 +pw4Xfam2) + getval("pwXechsel"); d.dutyoff = d1 + 4.0e-6; d.c1 = d.c1 + (!strcmp(dec.seq,"tppm")); d.c1 = d.c1 + ((!strcmp(dec.seq,"tppm")) && (dec.t.a > 0.0)); d.t1 = d2_ + tXechselinit + getval("rd") + getval("ad") + at; d.c2 = d.c2 + (!strcmp(dec.seq,"spinal")); d.c2 = d.c2 + ((!strcmp(dec.seq,"spinal")) && (dec.s.a > 0.0)); d.t2 = d2_ + tXechselinit + getval("rd") + getval("ad") + at; d = update_dutycycle(d); abort_dutycycle(d,10.0); // Set Phase Tables if (phase1 == 0) { settable(phf1Xmqmas,12,table1); settable(ph1Xfam2,6,table2); settable(ph2Xfam2,6,table3); settable(phfXechsel,96,table4); settable(phRec,48,table5); } else { settable(phf1Xmqmas,6,table6); settable(ph1Xfam2,6,table7); settable(ph2Xfam2,6,table8); settable(phfXechsel,48,table9); settable(phRec,24,table10); if (phase1 == 2) { tsadd(phf1Xmqmas,30,360); } } setreceiver(phRec); obsstepsize(1.0); // Begin Sequence xmtrphase(phf1Xmqmas); decphase(zero); obspower(getval("tpwr")); obspwrf(getval("aXmqmas")); obsunblank(); decunblank(); _unblank34(); delay(d1); sp1on(); delay(2.0e-6); sp1off(); delay(2.0e-6); // H Decoupler on Before MQMAS _dseqon(dec); // Two-Pulse MQMAS with DFS Conversion rgpulse(getval("pw1Xmqmas"),zero,0.0,0.0); xmtrphase(zero); txphase(ph1Xfam2); obspwrf(aXfam2); delay(d20); // X FAM2 Pulse loop(v4,v5); xmtron(); delay(pw1Xfam2); xmtroff(); txphase(ph2Xfam2); delay(pw2Xfam2); xmtron(); delay(pw3Xfam2); xmtroff(); txphase(ph2Xfam2); delay(pw4Xfam2); endloop(v5); // Tau Delay and Second Selective Echo Pulse xmtrphase(phfXechsel); obsblank(); obspower(getval("dbXechsel")); obspwrf(getval("aXechsel")); delay(3.0e-6); obsunblank(); delay(d21 + tXechsel); rgpulse(getval("pwXechsel"),zero,0.0,0.0); delay(d22); // Begin Acquisition obsblank(); _blank34(); delay(getval("rd")); startacq(getval("ad")); acquire(np, 1/sw); endacq(); _dseqoff(dec); obsunblank(); decunblank(); _unblank34(); }

pulsesequence() { int t1_counter; char CCLS[MAXSTR], /* C13 refocussing pulse in middle of t1 */ wtg3919[MAXSTR], f1180[MAXSTR]; /* Flag to start t1 @ halfdwell */ double timeCT=getval("timeCT"), tauxh, tau1, gzlvl3=getval("gzlvl3"), gzlvl4=getval("gzlvl4"), gt3=getval("gt3"), gt4=getval("gt4"), gstab=getval("gstab"), /* gradient recovery delay */ JNH = getval("JNH"), pwN = getval("pwN"), pwNlvl = getval("pwNlvl"), pwHs, tpwrs=0.0, compH=1.0, /* H1 90 degree pulse length at tpwrs */ sw1 = getval("sw1"), /* temporary Pbox parameters */ pwClvl = getval("pwClvl"), /* coarse power for C13 pulse */ pwC = getval("pwC"); /* C13 90 degree pulse length at pwClvl */ getstr("CCLS",CCLS); getstr("wtg3919",wtg3919); getstr("f1180",f1180); /* check validity of parameter range */ if((dm[A] == 'y' || dm[B] == 'y' || dm[C] == 'y' )) { text_error("incorrect Dec1 decoupler flags! "); psg_abort(1); } if((dm2[A] == 'y' || dm2[B] == 'y') ) { text_error("incorrect Dec2 decoupler flags! "); psg_abort(1); } if( dpwr2 > 50 ) { text_error("don't fry the probe, dpwr2 too large! "); psg_abort(1); } /* INITIALIZE VARIABLES */ if(wtg3919[0] != 'y') /* selective H20 one-lobe sinc pulse needs 1.69 */ { /* times more power than a square pulse */ pwHs = getval("pwHs"); compH = getval("compH"); } else pwHs = pw*2.385+7.0*rof1+d3*2.5; tauxh = ((JNH != 0.0) ? 1/(4*(JNH)) : 2.25e-3); setautocal(); /* activate auto-calibration flags */ if (autocal[0] == 'n') { if(wtg3919[0] != 'y') /* selective H20 one-lobe sinc pulse needs 1.69 */ { /* times more power than a square pulse */ if (pwHs > 1e-6) tpwrs = tpwr - 20.0*log10(pwHs/(compH*pw*1.69)); else tpwrs = 0.0; tpwrs = (int) (tpwrs); } } else /* if autocal = 'y'(yes), 'q'(quiet), r(read), or 's'(semi) */ { if(FIRST_FID) /* call Pbox */ { if(wtg3919[0] != 'y') H2Osinc = pbox_Rsh("H2Osinc", "sinc90", pwHs, 0.0, compH*pw, tpwr); } if (wtg3919[0] != 'y') { pwHs = H2Osinc.pw; tpwrs = H2Osinc.pwr-1.0; } /* 1dB correction applied */ } /* LOAD VARIABLES */ if(ix == 1) d2_init = d2; t1_counter = (int) ( (d2-d2_init)*sw1 + 0.5); /* Set up f1180 */ tau1 = d2; if((f1180[A] == 'y') && (ni > 1.0)) { tau1 += ( 1.0 / (2.0*sw1) ); if(tau1 < 0.2e-6) tau1 = 0.0; } tau1 = tau1/2.0; /* LOAD PHASE TABLES */ assign(one,v7); assign(three,v8); settable(t1, 4, phi1); settable(t2, 2, phi2); settable(t3, 8, phi3); settable(t4, 16, phi4); settable(t5, 8, rec); if ( phase1 == 2 ) tsadd(t2, 1, 4); if(t1_counter %2) /* calculate modification to phases based on */ { tsadd(t2,2,4); tsadd(t5,2,4); } /* current t1 values */ if(wtg3919[0] != 'y') { add(one,v7,v7); add(one,v8,v8); } /* sequence starts!! */ status(A); obspower(tpwr); dec2power(pwNlvl); decpower(pwClvl); decpwrf(4095.0); delay(d1); status(B); rgpulse(pw, zero, rof1, rof1); zgradpulse(0.3*gzlvl3,gt3); txphase(zero); dec2phase(zero); delay(tauxh-gt3); /* delay=1/4J(XH) */ sim3pulse(2*pw,0.0,2*pwN,t4,zero,zero,rof1,rof1); zgradpulse(0.3*gzlvl3,gt3); dec2phase(t2); delay(tauxh-gt3 ); /* delay=1/4J(XH) */ rgpulse(pw, t1, rof1, rof1); decphase(zero); txphase(t4); zgradpulse(gzlvl3,gt3); delay(gstab); dec2rgpulse(pwN, t2, rof1, rof1); /* CT EVOLUTION BEGINS */ dec2phase(t3); delay(timeCT -SAPS_DELAY -tau1); if (CCLS[A]=='y') { sim3pulse(0.0, 2.0*pwC, 2.0*pwN, zero, zero, zero, 0.0, 0.0); delay(timeCT -2.0*pw); rgpulse(2.0*pw, t4, 0.0, 0.0); } else { dec2rgpulse(2.0*pwN, zero, 0.0, 0.0); delay(timeCT -2.0*pwC); simpulse(2.0*pw, 2.0*pwC, t4, zero, 0.0, 0.0); } delay(tau1); /* CT EVOLUTION ENDS */ dec2rgpulse(pwN, t3, rof1, rof1); zgradpulse(gzlvl3,gt3); delay(gstab); rgpulse(pw, two, rof1, rof1); decrgpulse(pwC, zero, rof1, rof1); zgradpulse(gzlvl4,gt4); txphase(v7); dec2phase(zero); delay(tauxh -gt4 -pwHs -rof1 -2.0*pwC -2.0*rof1); if(wtg3919[0] == 'y') { rgpulse(pw*0.231,v7,rof1,rof1); delay(d3); rgpulse(pw*0.692,v7,rof1,rof1); delay(d3); rgpulse(pw*1.462,v7,rof1,rof1); delay(d3/2-pwN); dec2rgpulse(2*pwN, zero, rof1, rof1); txphase(v8); delay(d3/2-pwN); rgpulse(pw*1.462,v8,rof1,rof1); delay(d3); rgpulse(pw*0.692,v8,rof1,rof1); delay(d3); rgpulse(pw*0.231,v8,rof1,rof1); } else { obspower(tpwrs); shaped_pulse("H2Osinc", pwHs, v7, rof1, 0.0); obspower(tpwr); sim3pulse(2.0*pw, 0.0, 2.0*pwN, v8, zero, zero, 0.0, 0.0); obspower(tpwrs); shaped_pulse("H2Osinc", pwHs, v7, rof1, 0.0); obspower(tpwr); } zgradpulse(gzlvl4,gt4); delay(tauxh -gt4 -pwHs -rof1 -POWER_DELAY); dec2power(dpwr2); status(C); setreceiver(t5); }

pulsesequence() { /* DECLARE VARIABLES */ char autocal[MAXSTR], /* auto-calibration flag */ fsat[MAXSTR], fscuba[MAXSTR], f1180[MAXSTR], /* Flag to start t1 @ halfdwell */ f2180[MAXSTR], /* Flag to start t2 @ halfdwell */ c180_flg[MAXSTR], codecseq[MAXSTR], mess_flg[MAXSTR], ch_shp1[MAXSTR], /* shape for the 1st purge CHIRP */ ch_shp2[MAXSTR], /* shape for the 2nd purge CHIRP */ chshpi[MAXSTR]; /* shape for the INEPT CHIRPs */ int phase, phase2, t1_counter, /* used for states tppi in t1 */ t2_counter; /* used for states tppi in t2 */ double tau1, /* t1 delay */ tau2, /* t2 delay */ ni2, mix, /* mixing time in seconds */ pwC, /* PW90 for c nucleus @ pwClvl */ pwcodec, /* PW for C' nucleus @ dpwrco seduce dec */ tsatpwr, /* low level 1H trans.power for presat */ pwClvl, /* power level for 13C pulses on dec1 */ dpwrco, /* power level for C' seduce decoupling */ sw1, /* sweep width in f1 */ sw2, /* sweep width in f2 */ tofps, /* tof for presat */ dressed, /* decoupler resolution for seduce decoupling */ tpwrmess, /* power level for Messerlie purge */ dly_pg1, /* duration of first part of purge */ dly_wt, taua1, /* Delay for the first purge CHIRP */ taua2, /* Delay for the second purge CHIRP */ pwchirp1, /* duration of the 1st purge CHIRP */ pwchirp2, /* duration of the 2nd purge CHIRP */ d_me1, /* time difference between start of the sweep and the excitation of the methyl region automatically calculated by the program necessary parameter diff (see below) */ d_me2, /* time difference between start of the sweep and the excitation of the methyl region automatically calculated by the program necessary parameter diff (see below) */ dchrp1, /* power for the 1st purge CHIRP pulse, only lower limit is important (see above!) */ dchrp2, /* power for the 2nd purge CHIRP pulse, only lower limit is important (see above!) */ dmfchp1, /* dmf (1/90) for the 1st purge CHIRP pulse dmfchp1 = 1/time_step of chirp-pulse [time_step = pwchirp1/no. of points in the .DEC-shape] */ dmfchp2, /* dmf (1/90) for the 1st purge CHIRP pulse dmfchp2 = 1/time_step of chirp-pulse [time_step = pwchirp2/no. of points in the .DEC-shape] */ dres_chp, /* dres for the chirp pulse (must be set to 90, otherwise timing errors! ) */ diff1, /* shift differences between methyl region and start of sweep */ diff2, /* shift differences between methyl region and start of sweep */ rate1, /* sweep rate of the 1st purge CHIRP pulse frequency sweep/pwchirp1 */ rate2, /* sweep rate of the 2nd purge CHIRP pulse frequency sweep/pwchirp2 */ dchrpi, dmfchpi, pwchirpi, /* INEPT CHIRP duration */ ratei, diffi, tauf, d_mei, compC, /* C-13 RF calibration parameters */ gt0, gt1, gt2, gt3, gt4, gt5, gt8, gt9, gt10, gt11, gstab, gzlvl0, gzlvl1, gzlvl2, gzlvl3, gzlvl4, gzlvl5, gzlvl8, gzlvl9, gzlvl10, gzlvl11; /* variables commented out are already defined by the system */ /* LOAD VARIABLES */ getstr("autocal",autocal); getstr("fsat",fsat); getstr("f1180",f1180); getstr("f2180",f2180); getstr("fscuba",fscuba); getstr("c180_flg",c180_flg); getstr("mess_flg",mess_flg); tofps = getval("tofps"); mix = getval("mix"); pwC = getval("pwC"); tpwr = getval("tpwr"); tsatpwr = getval("tsatpwr"); pwClvl = getval("pwClvl"); dpwr = getval("dpwr"); dpwr2 = getval("dpwr2"); phase = (int) ( getval("phase") + 0.5); phase2 = (int) ( getval("phase2") + 0.5); sw1 = getval("sw1"); sw2 = getval("sw2"); ni2 = getval("ni2"); tpwrmess = getval("tpwrmess"); dly_pg1 = getval("dly_pg1"); dly_wt = getval("dly_wt"); taua1 = getval("taua1"); taua2 = getval("taua2"); rate1 = getval("rate1"); rate2 = getval("rate2"); diff1 = getval("diff1"); diff2 = getval("diff2"); diffi = getval("diffi"); ratei = getval("ratei"); tauf = getval("tauf"); gt0 = getval("gt0"); gt1 = getval("gt1"); gt2 = getval("gt2"); gt3 = getval("gt3"); gt4 = getval("gt4"); gt5 = getval("gt5"); gt8 = getval("gt8"); gt9 = getval("gt9"); gt10 = getval("gt10"); gt11 = getval("gt11"); gstab = getval("gstab"); gzlvl0 = getval("gzlvl0"); gzlvl1 = getval("gzlvl1"); gzlvl2 = getval("gzlvl2"); gzlvl3 = getval("gzlvl3"); gzlvl4 = getval("gzlvl4"); gzlvl5 = getval("gzlvl5"); gzlvl8 = getval("gzlvl8"); gzlvl9 = getval("gzlvl9"); gzlvl10 = getval("gzlvl10"); gzlvl11 = getval("gzlvl11"); if(autocal[0]=='n') { getstr("codecseq",codecseq); dressed = getval("dressed"); pwcodec = getval("pwcodec"); dpwrco = getval("dpwrco"); getstr("ch_shp1",ch_shp1); getstr("ch_shp2",ch_shp2); pwchirp1 = getval("pwchirp1"); pwchirp2 = getval("pwchirp2"); dchrp1 = getval("dchrp1"); dchrp2 = getval("dchrp2"); dmfchp1 = getval("dmfchp1"); dmfchp2 = getval("dmfchp2"); dres_chp = getval("dres_chp"); getstr("chshpi",chshpi); dchrpi = getval("dchrpi"); dmfchpi = getval("dmfchpi"); pwchirpi = getval("pwchirpi"); } else { strcpy(codecseq,"Psed_108p"); strcpy(ch_shp1,"Pwurst180_1"); strcpy(ch_shp2,"Pwurst180_2"); strcpy(chshpi,"Pwurst180i"); if (FIRST_FID) { compC = getval("compC"); codec = pbox(codecseq, CODEC, CODECps, dfrq, compC*pwC, pwClvl); chirp1 = pbox(ch_shp1, CHIRP1, CHIRPps, dfrq, compC*pwC, pwClvl); chirp2 = pbox(ch_shp2, CHIRP2, CHIRPps, dfrq, compC*pwC, pwClvl); chirpi = pbox(chshpi, CHIRPi, CHIRPps, dfrq, compC*pwC, pwClvl); } dpwrco = codec.pwr; pwcodec = 1.0/codec.dmf; dressed = codec.dres; dchrp1 = chirp1.pwr; dmfchp1 = chirp1.dmf; pwchirp1 = chirp1.pw; dres_chp = chirp1.dres; dchrp2 = chirp1.pwr; dmfchp2 = chirp2.dmf; pwchirp2 = chirp2.pw; dchrpi = chirpi.pwr; dmfchpi = chirpi.dmf; pwchirpi = chirpi.pw; } /* LOAD PHASE TABLE */ settable(t1,8,phi1); settable(t2,16,phi2); settable(t4,16,rec); settable(t5,4,phi5); settable(t6,2,phi6); settable(t7,4,phi7); /* CHECK VALIDITY OF PARAMETER RANGES */ if((dm[A] == 'y' || dm[B] == 'y' )) { printf("incorrect dec1 decoupler flags! "); psg_abort(1); } if((dm2[A] == 'y' || dm2[B] == 'y')) { printf("incorrect dec2 decoupler flags! Should be 'nnn' "); psg_abort(1); } if( tsatpwr > 6 ) { printf("TSATPWR too large !!! "); psg_abort(1); } if( dpwr > 50 ) { printf("don't fry the probe, DPWR too large! "); psg_abort(1); } if( dpwrco > 50 ) { printf("don't fry the probe, dpwrco too large! "); psg_abort(1); } if( dpwr2 > 46 ) { printf("don't fry the probe, DPWR2 too large! "); psg_abort(1); } if( pw > 200.0e-6 ) { printf("dont fry the probe, pw too high ! "); psg_abort(1); } if( pwC > 200.0e-6 ) { printf("dont fry the probe, pwC too high ! "); psg_abort(1); } if( pwcodec < 300.0e-6 ) { printf("dont fry the probe, pwcodec too high ! "); psg_abort(1); } if ( tpwrmess > 56 ) { printf("dont fry the probe, tpwrmess too high ! "); psg_abort(1); } if ( dly_pg1 > 0.010) { printf("dont fry the probe, dly_pg1 too long ! "); psg_abort(1); } if( gt0 > 15e-3 || gt1 > 15e-3 || gt2 > 15e-3 || gt3 > 15e-3 || gt4 > 15e-3 || gt5 > 15e-3 || gt8 > 15e-3 || gt9 > 15e-3 || gt10 > 15e-3 || gt11 > 15e-3 ) { printf("gti values < 15e-3\n"); psg_abort(1); } if( gzlvl3*gzlvl4 > 0.0 ) { printf("gt3 and gt4 must be of opposite sign \n"); printf("for optimal water suppression\n"); psg_abort(1); } if( dchrp1 > 60 ) { printf("don't fry the probe, dchrp1 too large! "); psg_abort(1); } if( dchrp2 > 60 ) { printf("don't fry the probe, dchrp2 too large! "); psg_abort(1); } if( pwchirp1 > 10.e-03 ) { printf("don't fry the probe, pwchirp1 too large! "); psg_abort(1); } if( pwchirp2 > 10.e-03 ) { printf("don't fry the probe, pwchirp2 too large! "); psg_abort(1); } d_me1 = diff1/rate1 ; d_me2 = diff2/rate2 ; if( d_me1 > 10.e-03 ) { printf("don't fry the probe, d_me1 too large \n"); printf(" (must be less than 10 msec)! "); psg_abort(1); } if( d_me2 > 10.e-03 ) { printf("don't fry the probe, d_me2 too large \n"); printf(" (must be less than 10 msec)! "); psg_abort(1); } if( d_me1 > pwchirp1 ) { printf("impossible; d_me1 > pwchirp1 ! "); psg_abort(1); } if( d_me2 > pwchirp2 ) { printf("impossible; d_me2 > pwchirp2 ! "); psg_abort(1); } if( dchrpi > 60 ) { printf("dont fry the probe, dchrpi too large\n"); psg_abort(1); } if(pwchirpi > 10.0e-3) { printf("don't fry the probe, pwchirpi too large! "); psg_abort(1); } d_mei = diffi/ratei; /* Phase incrementation for hypercomplex 2D data */ if (phase == 2) tsadd(t1,1,4); if (phase2 == 2) tsadd(t2,1,4); /* Set up f1180 tau1 = t1 */ tau1 = d2; if(f1180[A] == 'y') { tau1 += ( 1.0 / (2.0*sw1) - 4.0/PI*pw - 2.0e-6 ); } else tau1 = tau1 - 4.0/PI*pw - 2.0e-6; if(tau1 < 0.2e-6) tau1 = 2.0e-7; /* Set up f2180 tau2 = t2 */ tau2 = d3; if(f2180[A] == 'y') { tau2 += ( 1.0 / (2.0*sw2) - (4.0/PI)*pwC - 2.0*pw - PRG_START_DELAY - PRG_STOP_DELAY - 2.0*POWER_DELAY - 4.0e-6); } else tau2 = tau2 - ((4.0/PI)*pwC + 2.0*pw + PRG_START_DELAY + PRG_STOP_DELAY + 2.0*POWER_DELAY + 4.0e-6); if(tau2 < 0.2e-6) tau2 = 4.0e-7; tau2 = tau2/2.0; /* Calculate modifications to phases for States-TPPI acquisition */ if( ix == 1) d2_init = d2 ; t1_counter = (int) ( (d2-d2_init)*sw1 + 0.5 ); if(t1_counter % 2) { tsadd(t1,2,4); tsadd(t4,2,4); } if( ix == 1) d3_init = d3 ; t2_counter = (int) ( (d3-d3_init)*sw2 + 0.5 ); if(t2_counter % 2) { tsadd(t2,2,4); tsadd(t4,2,4); } /* BEGIN ACTUAL PULSE SEQUENCE */ status(A); delay(5.0e-6); obspower(tsatpwr); /* Set transmitter power for 1H presaturation */ decpower(pwClvl); /* Set Dec1 power for hard 13C pulses */ delay(5.0e-6); /* Presaturation Period */ if (mess_flg[A] == 'y') { obsoffset(tofps); obspower(tpwrmess); txphase(zero); rgpulse(dly_pg1,zero,2.0e-6,2.0e-6); txphase(one); rgpulse(dly_pg1/1.62,one,2.0e-6,2.0e-6); obspower(tsatpwr); } if (fsat[0] == 'y') { obsoffset(tofps); delay(2.0e-5); rgpulse(d1,zero,2.0e-6,2.0e-6); /* presat */ obspower(tpwr); /* Set transmitter power for hard 1H pulses */ delay(2.0e-5); if(fscuba[0] == 'y') { delay(2.2e-2); rgpulse(pw,zero,2.0e-6,0.0); rgpulse(2*pw,one,2.0e-6,0.0); rgpulse(pw,zero,2.0e-6,0.0); delay(2.2e-2); } } else { delay(d1); } obspower(tpwr); /* Set transmitter power for hard 1H pulses */ obsoffset(tof); decphase(zero); /* Begin Pulses */ status(B); rcvroff(); delay(10.0e-6); rgpulse(pw,t6,4.0e-6,0.0); /* 90 deg 1H pulse */ txphase(zero); decphase(zero); delay(2.0e-6); zgradpulse(gzlvl8,gt8); delay(gstab); delay(taua1 - gt8 - gstab -2.0e-6 - POWER_DELAY - 4.0e-6 - PRG_START_DELAY - d_me1); /* 1st purge CHIRP inversion on */ decpower(dchrp1); /* Set power for 1st purge CHIRP inversion */ delay(4.0e-6); decprgon(ch_shp1,1.0/dmfchp1,dres_chp); decon(); delay(d_me1); rgpulse(2*pw,zero,0.0,0.0); /* 1H inversion pulse */ delay(pwchirp1 - d_me1 - 2*pw); decoff(); decprgoff(); /* chirp inversion off */ delay(2.0e-6); zgradpulse(gzlvl8,gt8); delay(gstab); delay(taua1 + 2*pw - (pwchirp1 - d_me1) - PRG_STOP_DELAY - gt8 - gstab -2.0e-6); rgpulse(pw,zero,0.0,0.0); txphase(t7); delay(2.0e-6); zgradpulse(gzlvl9,gt9); delay(2.0*gstab); rgpulse(pw,t7,0.0,0.0); /* PHASE t7 = 2(x),2(-x)*/ delay(2.0e-6); zgradpulse(gzlvl11,gt11); delay(gstab); decphase(zero); txphase(zero); delay(taua2 - gt11 - gstab -2.0e-6 - POWER_DELAY - 4.0e-6 - PRG_START_DELAY - d_me2); /* Second chirp inversion on */ decpower(dchrp2); /* Set power for chirp inversion */ delay(4.0e-6); decprgon(ch_shp2,1.0/dmfchp2,dres_chp); decon(); delay(d_me2); rgpulse(2*pw,zero,0.0,0.0); /* 1H inversion pulse */ delay(pwchirp2 - d_me2 - 2*pw); decoff(); decprgoff(); /* Second purge CHIRP off */ delay(2.0e-6); zgradpulse(gzlvl11,gt11); delay(gstab); txphase(zero); delay(taua2 + 2*pw - (pwchirp2 - d_me2) - PRG_STOP_DELAY - gt11 - gstab -2.0e-6 ); rgpulse(pw,zero,0.0,0.0); delay(2.0e-6); zgradpulse(gzlvl10,gt10); delay(2.0*gstab); rgpulse(pw,t1,4.0e-6,0.0); delay(tau1); rgpulse(pw,zero,2.0e-6,0.0); delay(mix - 10.0e-3); delay(2.0e-6); zgradpulse(gzlvl0,gt0); decpower(pwClvl); /* Set power for hard pulses */ delay(4.0e-6); decrgpulse(pwC,zero,0.0,0.0); delay(2.0e-6); zgradpulse(gzlvl1,gt1); delay(2.0e-6); decphase(zero); delay(10.0e-3 - gt1 - gt0 - 8.0e-6); rgpulse(pw,zero,0.0,0.0); delay(2.0e-6); zgradpulse(gzlvl2,gt2); delay(gstab); decphase(zero); delay(tauf - gt2 - gstab -2.0e-6 - POWER_DELAY - 4.0e-6 - PRG_START_DELAY - d_mei); /* INEPT CHIRP inversion on */ decpower(dchrpi); /* Set power for chirp inversion */ delay(4.0e-6); decprgon(chshpi,1.0/dmfchpi,dres_chp); decon(); delay(d_mei); rgpulse(2*pw,zero,0.0,0.0); /* 1H inversion pulse */ delay(pwchirpi - d_mei - 2*pw); decoff(); decprgoff(); /* chirp inversion off */ delay(2.0e-6); zgradpulse(gzlvl2,gt2); delay(gstab); txphase(one); delay(tauf + 2*pw - (pwchirpi - d_mei) - PRG_STOP_DELAY - gt2 - gstab -2.0e-6 ); rgpulse(pw,one,0.0,0.0); txphase(zero); decphase(t2); decpower(pwClvl); /* Set power for C13 hard pulse */ delay(2.0e-6); zgradpulse(gzlvl3,gt3); delay(200.0e-6); decrgpulse(pwC,t2,0.0,0.0); decphase(zero); if( c180_flg[A] == 'n' ) { delay(2.0e-6); /* CO decoupling on */ decpower(dpwrco); decprgon(codecseq,pwcodec,dressed); decon(); /* CO decoupling on */ delay(tau2); rgpulse(2*pw,zero,0.0,0.0); delay(tau2); /* CO decoupling off */ decoff(); decprgoff(); decpower(pwClvl); /* CO decoupling off */ delay(2.0e-6); } else simpulse(2*pw,2*pwC,zero,zero,2.0e-6,2.0e-6); decrgpulse(pwC,zero,0.0,0.0); delay(2.0e-6); zgradpulse(gzlvl4,gt4); delay(200.0e-6); rgpulse(pw,t5,2.0e-6,0.0); txphase(zero); delay(2.0e-6); zgradpulse(gzlvl5,gt5); delay(gstab); decphase(zero); delay(tauf - gt5 - gstab -2.0e-6 - POWER_DELAY - 4.0e-6 - PRG_START_DELAY - d_mei); /* 2nd INEPT CHIRP inversion on */ decpower(dchrpi); /* Set power for chirp inversion */ delay(4.0e-6); decprgon(chshpi,1.0/dmfchpi,dres_chp); decon(); delay(d_mei); rgpulse(2*pw,zero,0.0,0.0); /* 1H inversion pulse */ delay(pwchirpi - d_mei - 2*pw); decoff(); decprgoff(); /* chirp inversion off */ delay(2.0e-6); zgradpulse(gzlvl5,gt5); delay(gstab); decpower(dpwr); /* Set power for decoupling */ txphase(t5); delay(tauf + 2*pw - (pwchirpi - d_mei) - PRG_STOP_DELAY - gt5 - gstab -2.0e-6 - 2*POWER_DELAY); rgpulse(pw,t5,0.0,0.0); /* BEGIN ACQUISITION */ status(C); setreceiver(t4); }

pulsesequence() { /* DECLARE VARIABLES */ char autocal[MAXSTR], /* auto-calibration flag */ fsat[MAXSTR], fscuba[MAXSTR], f1180[MAXSTR], /* Flag to start t1 @ halfdwell */ mess_flg[MAXSTR], /* water purging */ ar180a[MAXSTR], /* waveform shape for aromatic 180 pulse with C transmitter at dof */ cb180b[MAXSTR], /* waveform shape for aliphatic 180 pulse with C transmitter at dofar */ ar180b[MAXSTR]; /* waveform shape for aromatic 180 pulse with C transmitter at dofar */ int phase, ni, t1_counter; /* used for states tppi in t1 */ double tau1, /* t1 delay */ taua, /* ~ 1/4JCbHb = 1.7 ms */ taub, /* ~ 1/4JCgCd = 2.7 ms */ tauc, /* ~ 1/4JCgCd = 2.1 ms */ taud, /* ~ 1/4JCdHd = 1.5 ms */ taue, /* = 1/4JCbHb = 1.8 ms */ tauf, /* 2(tauc-tauf) ~ 1/2JCdHd = 3.1 ms */ TCb, /* carbon constant time period for recording the Cb chemical shifts */ dly_pg1, /* delay for water purging */ pwar180a, /* 180 aro pulse at d_ar180a and dof */ pwcb180b, /* 180 cb pulse at d_cb180b and dofar */ pwC, /* 90 c pulse at pwClvl */ pwsel90, /* 90 c pulse at d_sel90 */ pwar180b, /* 180 c pulse at d_ar180b */ compC, /* C-13 RF calibration parameters */ compH, d_ar180a, d_cb180b, d_sel90, d_ar180b, dofar, tsatpwr, /* low level 1H trans.power for presat */ tpwrmess, /* power level for water purging */ tpwrml, /* power level for 1H decoupling */ pwmlev, /* 90 pulse at tpwrml */ pwClvl, /* power level for high power 13C pulses on dec1 */ sw1, /* sweep width in f1 */ at, gp11, /* gap between 90-90 for selective 180 of Cb */ fab, /* chemical shift difference of Ca-Cb (Hz) */ gt0, gt1, gt2, gt3, gt4, gt5, gt6, gt7, gstab, gzlvl0, gzlvl1, gzlvl2, gzlvl3, gzlvl4, gzlvl5, gzlvl6, gzlvl7; /* variables commented out are already defined by the system */ /* LOAD VARIABLES */ getstr("autocal",autocal); getstr("fsat",fsat); getstr("f1180",f1180); getstr("fscuba",fscuba); getstr("mess_flg",mess_flg); taua = getval("taua"); taub = getval("taub"); tauc = getval("tauc"); taud = getval("taud"); taue = getval("taue"); tauf = getval("tauf"); TCb = getval("TCb"); pwC = getval("pwC"); dofar = getval("dofar"); dly_pg1 = getval("dly_pg1"); tpwr = getval("tpwr"); tsatpwr = getval("tsatpwr"); tpwrmess = getval("tpwrmess"); tpwrml = getval("tpwrml"); pwClvl = getval("pwClvl"); dpwr = getval("dpwr"); phase = (int) ( getval("phase") + 0.5); sw1 = getval("sw1"); ni = getval("ni"); at = getval("at"); fab = getval("fab"); if(autocal[0]=='n') { getstr("ar180a",ar180a); getstr("ar180b",ar180b); getstr("cb180b",cb180b); pwar180a = getval("pwar180a"); pwar180b = getval("pwar180b"); pwcb180b = getval("pwcb180b"); pwsel90 = getval("pwsel90"); d_ar180a = getval("d_ar180a"); d_cb180b = getval("d_cb180b"); d_ar180b = getval("d_ar180b"); d_sel90 = getval("d_sel90"); pwmlev = getval("pwmlev"); } else { strcpy(ar180a,"Pg3_off_cb180a"); strcpy(ar180b,"Pg3_off_cb180b"); strcpy(cb180b,"Pg3_on"); if (FIRST_FID) { compC = getval("compC"); compH = getval("compH"); sel90 = pbox("cal", SEL90, "", dfrq, compC*pwC, pwClvl); ar_180a = pbox(ar180a, AR180a, CB180ps, dfrq, compC*pwC, pwClvl); ar_180b = pbox(ar180b, AR180b, CB180ps, dfrq, compC*pwC, pwClvl); cb_180b = pbox(cb180b, CB180b, CB180ps, dfrq, compC*pwC, pwClvl); w16 = pbox_dec("cal", "WALTZ16", tpwrml, sfrq, compH*pw, tpwr); } pwsel90 = sel90.pw; d_sel90 = sel90.pwr; pwar180a = ar_180a.pw; d_ar180a = ar_180a.pwr; pwar180b = ar_180b.pw; d_ar180b = ar_180b.pwr; pwcb180b = cb_180b.pw; d_cb180b = cb_180b.pwr; pwmlev = 1.0/w16.dmf; } gt0 = getval("gt0"); gt1 = getval("gt1"); gt2 = getval("gt2"); gt3 = getval("gt3"); gt4 = getval("gt4"); gt5 = getval("gt5"); gt6 = getval("gt6"); gstab = getval("gstab"); gt7 = getval("gt7"); gzlvl0 = getval("gzlvl0"); gzlvl1 = getval("gzlvl1"); gzlvl2 = getval("gzlvl2"); gzlvl3 = getval("gzlvl3"); gzlvl4 = getval("gzlvl4"); gzlvl5 = getval("gzlvl5"); gzlvl6 = getval("gzlvl6"); gzlvl7 = getval("gzlvl7"); /* LOAD PHASE TABLE */ settable(t1,1,phi1); settable(t2,1,phi2); settable(t3,4,phi3); settable(t4,8,phi4); settable(t5,1,phi5); settable(t6,8,rec); /* CHECK VALIDITY OF PARAMETER RANGES */ if( 0.5*ni*1/(sw1) > TCb - 2*POWER_DELAY - WFG_START_DELAY - pwar180a - WFG_STOP_DELAY) { printf(" ni is too big\n"); psg_abort(1); } if((dm[A] == 'y' || dm[B] == 'y' )) { printf("incorrect dec1 decoupler flags! "); psg_abort(1); } if(dm2[A] == 'y' || dm2[B] == 'y' || dm2[C] == 'y' ) { printf("incorrect dec2 decoupler flags! "); psg_abort(1); } if( tsatpwr > 6 ) { printf("TSATPWR too large !!! "); psg_abort(1); } if( tpwrml > 53 ) { printf("tpwrml too large !!! "); psg_abort(1); } if( tpwrmess > 56 ) { printf("tpwrmess too large !!! "); psg_abort(1); } if( dpwr > 50 ) { printf("don't fry the probe, DPWR too large! "); psg_abort(1); } if( dpwr2 > 50 ) { printf("don't fry the probe, DPWR2 too large! "); psg_abort(1); } if( pwClvl > 63 ) { printf("don't fry the probe, DHPWR too large! "); psg_abort(1); } if( pw > 20.0e-6 ) { printf("dont fry the probe, pw too high ! "); psg_abort(1); } if( pwcb180b > 500.0e-6 ) { printf("dont fry the probe, pwcb180b too high ! "); psg_abort(1); } if( pwar180a > 500.0e-6 ) { printf("dont fry the probe, pwar180a too high ! "); psg_abort(1); } if (pwar180b > 500.0e-6) { printf("dont fry the probe, pwar180b too long !"); psg_abort(1); } if (pwsel90 > 100.0e-6) { printf("dont fry the probe, pwsel90 too long !"); psg_abort(1); } if(d_ar180a > 60) { printf("dont fry the probe, d_ar180a too high !"); psg_abort(1); } if(d_cb180b > 60) { printf("dont fry the probe, d_cb180b too high !"); psg_abort(1); } if (d_ar180b > 60) { printf("dont fry the probe, d_ar180b too high ! "); psg_abort(1); } if (d_sel90 > 50) { printf("dont fry the probe, d_sel90 too high ! "); psg_abort(1); } if( gt0 > 15e-3 || gt1 > 15e-3 || gt2 > 15e-3 || gt3 > 15e-3 || gt4 > 15e-3 || gt5 > 15e-3 || gt6 > 15e-3 || gt7 > 15e-3) { printf("gradients on for too long. Must be < 15e-3 \n"); psg_abort(1); } if( fabs(gzlvl0) > 30000 || fabs(gzlvl1) > 30000 || fabs(gzlvl2) > 30000 ||fabs(gzlvl3) > 30000 || fabs(gzlvl4) > 30000 || fabs(gzlvl5) > 30000 ||fabs(gzlvl6) > 30000 || fabs(gzlvl7) > 30000) { printf("too strong gradient"); psg_abort(1); } if( 2*TCb - taue > 0.1 ) { printf("dont fry the probe, too long TCb"); psg_abort(1); } if( at > 0.1 && (dm[C]=='y' || dm2[C]=='y')) { printf("dont fry the probe, too long at with decoupling"); psg_abort(1); } if( pwC > 30.0e-6) { printf("dont fry the probe, too long pwC"); psg_abort(1); } if( dly_pg1 > 10.0e-3) { printf("dont fry the probe, too long dly_pg1"); psg_abort(1); } /* Phase incrementation for hypercomplex 2D data */ if (phase == 2) tsadd(t2,1,4); /* Calculate modifications to phases for States-TPPI acquisition */ if( ix == 1) d2_init = d2 ; t1_counter = (int) ( (d2-d2_init)*sw1 + 0.5 ); if(t1_counter % 2) { tsadd(t2,2,4); tsadd(t6,2,4); } /* Set up f1180 tau1 = t1 */ tau1 = d2; if(f1180[A] == 'y') { tau1 += ( 1.0 / (2.0*sw1) ); } tau1 = tau1/2.0; /* 90-90 pulse for selective 180 of Cb but not Ca */ gp11 = 1/(2*fab) - 4/PI*pwsel90; if (gp11 < 0.0) { printf("gap of 90-90 negative, check fab and pwsel90"); psg_abort(1); } /* BEGIN ACTUAL PULSE SEQUENCE */ /* Receiver off time */ status(A); decoffset(dof); obspower(tsatpwr); /* Set transmitter power for 1H presaturation */ decpower(pwClvl); /* Set Dec1 power for hard 13C pulses */ dec2power(dpwr2); /* Set Dec2 power for 15N decoupling */ /* Presaturation Period */ if(mess_flg[A] == 'y') { obspower(tpwrmess); rgpulse(dly_pg1,zero,20.0e-6,20.0e-6); rgpulse(dly_pg1/1.62,one,20.0e-6,20.0e-6); obspower(tsatpwr); } if (fsat[0] == 'y') { delay(2.0e-5); rgpulse(d1,zero,20.0e-6,20.0e-6); obspower(tpwr); /* Set transmitter power for hard 1H pulses */ delay(2.0e-5); if(fscuba[0] == 'y') { delay(2.2e-2); rgpulse(pw,zero,2.0e-6,0.0); rgpulse(2*pw,one,2.0e-6,0.0); rgpulse(pw,zero,2.0e-6,0.0); delay(2.2e-2); } } else { delay(d1); } obspower(tpwr); /* Set transmitter power for hard 1H pulses */ txphase(zero); dec2phase(zero); decphase(zero); delay(1.0e-5); /* Begin Pulses */ rcvroff(); delay(10.0e-6); /* first ensure that magnetization does infact start on H and not C */ decrgpulse(pwC,zero,2.0e-6,2.0e-6); delay(2.0e-6); zgradpulse(gzlvl0,gt0); delay(gstab); /* this is the real start */ rgpulse(pw,zero,0.0,0.0); /* 90 deg 1H pulse */ delay(2.0e-6); zgradpulse(gzlvl1,gt1); delay(2.0e-6); delay(taua - gt1 - 4.0e-6); /* taua <= 1/4JCH */ simpulse(2*pw,2*pwC,zero,zero,0.0,0.0); txphase(t1); delay(2.0e-6); zgradpulse(gzlvl1,gt1); delay(2.0e-6); delay(taua - gt1 - 4.0e-6); rgpulse(pw,t1,0.0,0.0); txphase(zero); delay(2.0e-6); zgradpulse(gzlvl2,gt2); delay(gstab); decphase(t2); decpower(d_sel90); decrgpulse(pwsel90,t2,2.0e-6,0.0); decphase(zero); decpower(d_ar180a); decshaped_pulse(ar180a,pwar180a,zero,2.0e-6,0.0); /* bs effect */ delay(taue - POWER_DELAY - 2.0e-6 - WFG_START_DELAY - pwar180a - WFG_STOP_DELAY - POWER_DELAY - PRG_START_DELAY); /* H decoupling on */ obspower(tpwrml); obsprgon("waltz16",pwmlev,90.0); xmtron(); /* TURN ME OFF DONT FORGET */ /* Hldecoupling on */ delay(TCb + tau1 - taue - POWER_DELAY - 2.0e-6); decphase(t3); decpower(d_sel90); decrgpulse(pwsel90,t3,2.0e-6,0.0); delay(gp11); decrgpulse(pwsel90,t3,0.0,0.0); decphase(zero); decpower(d_ar180a); decshaped_pulse(ar180a,pwar180a,zero,2.0e-6,0.0); delay(TCb - tau1 - POWER_DELAY - WFG_START_DELAY - 2.0e-6 - pwar180a - WFG_STOP_DELAY - POWER_DELAY - 2.0e-6); decphase(zero); decpower(d_sel90); decrgpulse(pwsel90,zero,2.0e-6,0.0); /* H decoupling off */ xmtroff(); obsprgoff(); obspower(tpwr); /* H decoupling off */ decoffset(dofar); delay(2.0e-6); zgradpulse(gzlvl3,gt3); delay(gstab); decphase(t4); decpower(d_sel90); decrgpulse(pwsel90,t4,2.0e-6,0.0); decphase(zero); decpower(d_cb180b); decshaped_pulse(cb180b,pwcb180b,zero,2.0e-6,0.0); /* B.S. */ delay(2.0e-6); zgradpulse(gzlvl4,gt4); delay(2.0e-6); delay(taub - POWER_DELAY - WFG_START_DELAY - 2.0e-6 - pwcb180b - WFG_STOP_DELAY - gt4 - 4.0e-6 - POWER_DELAY - WFG_START_DELAY - 2.0e-6); decphase(zero); decpower(d_ar180b); decshaped_pulse(ar180b,pwar180b,zero,2.0e-6,0.0); decpower(d_cb180b); decshaped_pulse(cb180b,pwcb180b,zero,2.0e-6,0.0); delay(2.0e-6); zgradpulse(gzlvl4,gt4); delay(2.0e-6); delay(taub - WFG_STOP_DELAY - POWER_DELAY - WFG_START_DELAY - 2.0e-6 - pwcb180b - WFG_STOP_DELAY - gt4 - 4.0e-6 - POWER_DELAY - 2.0e-6); decpower(d_sel90); decrgpulse(pwsel90,zero,2.0e-6,0.0); delay(2.0e-6); zgradpulse(gzlvl5,gt5); delay(100.0e-6); delay(tauc - POWER_DELAY - gt5 - 102.0e-6 - 2.0e-6); decphase(t5); decpower(pwClvl); decrgpulse(2*pwC,t5,2.0e-6,0.0); delay(2.0e-6); zgradpulse(gzlvl5,gt5); delay(100.0e-6); txphase(zero); delay(tauf - gt5 - 102.0e-6); rgpulse(2*pw,zero,0.0,0.0); delay(tauc - tauf - 2*pw - POWER_DELAY - 2.0e-6); decphase(zero); decpower(d_sel90); decrgpulse(pwsel90,zero,2.0e-6,0.0); txphase(zero); delay(2.0e-6); zgradpulse(gzlvl6,gt6); delay(gstab); rgpulse(pw,zero,0.0,0.0); delay(2.0e-6); zgradpulse(gzlvl7,gt7); delay(2.0e-6); delay(taud - gt7 - 4.0e-6 - POWER_DELAY - 2.0e-6); decphase(zero); decpower(pwClvl); simpulse(2*pw,2*pwC,zero,zero,2.0e-6,0.0); delay(2.0e-6); zgradpulse(gzlvl7,gt7); delay(2.0e-6); delay(taud - gt7 - 4.0e-6 - 2*POWER_DELAY); decpower(dpwr); /* Set power for decoupling */ dec2power(dpwr2); /* Set power for decoupling */ rgpulse(pw,zero,0.0,0.0); /* rcvron(); */ /* Turn on receiver to warm up before acq */ /* BEGIN ACQUISITION */ status(C); setreceiver(t6); }

void pulsesequence() { char shname1[MAXSTR], f1180[MAXSTR], f2180[MAXSTR], CT_flg[MAXSTR], n15_flg[MAXSTR]; int icosel, t1_counter, t2_counter, ni2 = getval("ni2"), phase; double d2_init=0.0, d3_init=0.0, pwS1,pwS2,pwS3,pwS4,pwS5,pwS6,pwS7,max, kappa, lambda = getval("lambda"), gzlvl1 = getval("gzlvl1"), gzlvl2 = getval("gzlvl2"), gzlvl3 = getval("gzlvl3"), gzlvl4 = getval("gzlvl4"), gzlvl5 = getval("gzlvl5"), gzlvl6 = getval("gzlvl6"), gt1 = getval("gt1"), gt3 = getval("gt3"), gt4 = getval("gt4"), gt5 = getval("gt5"), gt6 = getval("gt6"), gstab = getval("gstab"), tpwrsf = getval("tpwrsf"), shlvl1, shpw1 = getval("shpw1"), pwClvl = getval("pwClvl"), pwNlvl = getval("pwNlvl"), pwN = getval("pwN"), dpwr2 = getval("dpwr2"), d2 = getval("d2"), shbw = getval("shbw"), shofs = getval("shofs")-4.77, scale = getval("scale"), sw1 = getval("sw1"), timeTN = getval("timeTN"), timeTN1, Delta, t2a,t2b,halfT2,ctdelay, tauNCO = getval("tauNCO"), CTdelay = getval("CTdelay"), tauC = getval("tauC"), tau1, tau2, taunh = getval("taunh"); getstr("shname1", shname1); getstr("CT_flg",CT_flg); getstr("n15_flg",n15_flg); getstr("f1180",f1180); getstr("f2180",f2180); phase = (int) (getval("phase") + 0.5); settable(t1,2,phi1); settable(t3,1,phi3); settable(t4,4,phi4); settable(t5,1,phi5); settable(t14,4,phi14); settable(t24,4,phi24); /* INITIALIZE VARIABLES */ timeTN1= timeTN-tauC; Delta = timeTN-tauC-tauNCO; //shpw1 = pw*8.0; shlvl1=tpwr; pwS1 = c13pulsepw("ca", "co", "square", 90.0); pwS2 = c13pulsepw("ca", "co", "square", 180.0); pwS3 = c13pulsepw("co", "ca", "sinc", 180.0); pwS7 = c13pulsepw("co", "ca", "sinc", 90.0); pwS4 = h_shapedpw("eburp2",shbw,shofs,zero, 0.0, 0.0); pwS6 = h_shapedpw("reburp",shbw,shofs,zero, 0.0, 0.0); pwS5 = h_shapedpw("pc9f",shbw,shofs,zero, 2.0e-6, 0.0); if (CT_flg[0] == 'y') { if ( ni*1/(sw1)/2.0 > (CTdelay*0.5-gt3-1.0e-4)) { printf(" ni is too big. Make ni equal to %d or less.\n", ((int)((CTdelay*0.5-gt3-1.0e-4)*2.0*sw1))); psg_abort(1);} } if (phase == 1) ; if (phase == 2) {tsadd(t1,1,4);} if ( phase2 == 2 ) { tsadd ( t3,2,4 ); tsadd ( t5,2,4 ); icosel = +1; } else icosel = -1; /* Set up f1180 */ tau1 = d2; if((f1180[A] == 'y') && (ni > 1.0)) { tau1 += ( 1.0 / (2.0*sw1) ); if(tau1 < 0.2e-6) tau1 = 0.0; } /* Set up f2180 */ tau2 = d3; if((f2180[A] == 'y') && (ni2 > 1.0)) { tau2 += ( 1.0 / (2.0*sw2) ); if(tau2 < 0.2e-6) tau2 = 0.0; } /************************************************************/ if( ix == 1) d2_init = d2; t1_counter = (int) ( (d2-d2_init)*sw1 + 0.5 ); if(t1_counter % 2) { tsadd(t1,2,4); tsadd(t14,2,4); tsadd(t24,2,4); } if( ix == 1) d3_init = d3; t2_counter = (int) ( (d3-d3_init)*sw2 + 0.5 ); if(t2_counter % 2) { tsadd(t4,2,4); tsadd(t14,2,4); tsadd(t24,2,4); } /************************************************************/ /* Set up CONSTANT/SEMI-CONSTANT time evolution in N15 */ /************************************************************/ ctdelay = timeTN1-gt1-1.0e-4; // ctdelay = timeTN1-gt1-1.0e-4-2.0*GRADIENT_DELAY-4*POWER_DELAY-4*PWRF_DELAY-(4/PI)*pwN; if (ni2 > 1) { halfT2 = 0.5*(ni2-1)/sw2; t2b = (double) t2_counter*((halfT2 - ctdelay)/((double)(ni2-1))); if( ix==1 && halfT2 - timeTN > 0 ) printf("SCT mode on, max ni2=%g\n",timeTN*sw2*2+1); if(t2b < 0.0) t2b = 0.0; t2a = ctdelay - tau2*0.5 + t2b; if(t2a < 0.2e-6) t2a = 0.0; } else { t2b = 0.0; t2a = ctdelay - tau2*0.5; } /************************************************************/ status(A); rcvroff(); decpower(pwClvl); decoffset(dof); dec2power(pwNlvl); dec2offset(dof2); obspwrf(tpwrsf); decpwrf(4095.0); obsoffset(tof); set_c13offset("co"); dec2rgpulse(pwN*2.0,zero,0.0,0.0); zgradpulse(gzlvl4, gt4); delay(1.0e-4); delay(d1-gt4); lk_hold(); h_shapedpulse("pc9f",shbw,shofs,zero, 2.0e-6, 0.0); delay(lambda-pwS5*0.5-pwS6*0.4); h_sim3shapedpulse("reburp",shbw,shofs,0.0,2.0*pwN, one, zero, zero, 0.0, 0.0); delay(lambda-pwS5*0.5-pwS6*0.4); if(n15_flg[0]=='y') h_shapedpulse("pc9f_",shbw,shofs,three, 0.0, 0.0); else h_shapedpulse("pc9f_",shbw,shofs,one, 0.0, 0.0); zgradpulse(gzlvl4, gt4*4.0); delay(1.0e-4); obspower(shlvl1); /**************************************************************************/ /* xxxxxxxxxxxxxxxxxxxxxx N-> CA transfer xxxxxxxxxxxxxxxxxx */ /**************************************************************************/ dec2rgpulse(pwN,zero,0.0,0.0); delay(timeTN1); sim3_c13pulse("", "ca", "co", "square", "", 0.0, 180.0, 2.0*pwN, zero, zero, zero, 2.0e-6, 2.0e-6); delay(Delta); c13pulse("co", "ca", "sinc", 180.0, zero, 0.0, 0.0); delay(timeTN1-Delta-pwS3+pwN*4.0/3.0); c13pulse("co", "ca", "sinc", 90.0, zero, 0.0, 0.0); delay(tauC); c13pulse("ca", "co", "square", 180.0, zero, 2.0e-6, 2.0e-6); sim3_c13pulse("", "co", "ca", "sinc", "", 0.0, 180.0, 2.0*pwN, zero, zero, zero, 2.0e-6, 2.0e-6); delay(tauC); c13pulse("ca", "co", "square", 180.0, zero, 2.0e-6, 2.0e-6); c13pulse("co", "ca", "sinc", 90.0, one, 0.0, 0.0); dec2rgpulse(pwN,one,0.0,0.0); /**************************************************************************/ /* xxxxxxxxxxxxxxxxxxxxxx 13CA EVOLUTION xxxxxxxxxxxxxxxxxx */ /**************************************************************************/ set_c13offset("ca"); c13pulse("ca", "co", "square", 90.0, t1, 2.0e-6, 0.0); if(CT_flg[0]=='y') { delay(tau1*0.5); sim3_c13pulse(shname1, "co", "ca", "sinc", "", shpw1, 180.0, 2.0*pwN, zero, zero, zero, 2.0e-6, 2.0e-6); zgradpulse(gzlvl3, gt3); delay(1.0e-4); delay(CTdelay*0.5-gt3-1.0e-4); c13pulse("cab", "co", "square", 180.0, zero, 2.0e-6, 2.0e-6); delay(CTdelay*0.5-gt3-1.0e-4-tau1*0.5); sim3_c13pulse(shname1, "co", "ca", "sinc", "", shpw1, 180.0, 2.0*pwN, zero, zero, zero, 2.0e-6, 2.0e-6); zgradpulse(gzlvl3, gt3); delay(1.0e-4); } else { delay(tau1*0.5); sim3_c13pulse(shname1, "co", "ca", "sinc", "", shpw1, 180.0, 2.0*pwN, zero, zero, zero, 2.0e-6, 2.0e-6); zgradpulse(gzlvl3, gt3); delay(1.0e-4); delay(tau1*0.5); c13pulse("ca", "co", "square", 180.0, zero, 2.0e-6, 2.0e-6); sim3_c13pulse(shname1, "co", "ca", "sinc", "", shpw1, 180.0, 2.0*pwN, zero, zero, zero, 2.0e-6, 2.0e-6); zgradpulse(gzlvl3, gt3); delay(1.0e-4); } c13pulse("ca", "co", "square", 90.0, zero, 0.0, 0.0); set_c13offset("co"); /**************************************************************************/ /* xxxxxxxxxxxxxxxxxxxxxx N-> CA back transfer xxxxxxxxxxxxxxx */ /**************************************************************************/ obspower(shlvl1); dec2rgpulse(pwN,t4,0.0,0.0); c13pulse("co", "ca", "sinc", 90.0, one, 0.0, 0.0); c13pulse("ca", "co", "square", 180.0, zero, 2.0e-6, 2.0e-6); delay(tauC); sim3_c13pulse("", "co", "ca", "sinc", "", 0.0, 180.0, 2.0*pwN, zero, zero, zero, 2.0e-6, 2.0e-6); c13pulse("ca", "co", "square", 180.0, zero, 2.0e-6, 2.0e-6); delay(tauC); c13pulse("co", "ca", "sinc", 90.0, zero, 0.0, 0.0); // delay(timeTN1-Delta+tau2*0.5-pwS2-pwS3); delay(timeTN1-Delta+tau2*0.5-pwS2-pwS3-2.0*GRADIENT_DELAY+4*POWER_DELAY+4*PWRF_DELAY); c13pulse("co", "ca", "sinc", 180.0, zero, 0.0, 0.0); delay(Delta); c13pulse("ca", "co", "square", 180.0, zero, 0.0, 0.0); delay (t2b); dec2rgpulse (2.0*pwN, zero, 0.0, 0.0); zgradpulse(gzlvl1, gt1); delay(1.0e-4); delay (t2a); /**************************************************************************/ /** gradient-selected TROSY sequence *********/ /**************************************************************************/ delay(gt1/10.0+1.0e-4); h_shapedpulse("eburp2_",shbw,shofs,t3, 2.0e-6, 0.0); zgradpulse(gzlvl5, gt5); delay(lambda-pwS6*0.5-pwS4*scale- gt5); h_sim3shapedpulse("reburp",shbw,shofs,0.0,2.0*pwN, zero, zero, zero, 0.0, 0.0); zgradpulse(gzlvl5, gt5); delay(lambda-pwS6*0.5-pwS4*scale- gt5); h_shapedpulse("eburp2",shbw,shofs,zero, 0.0, 0.0); delay(gt1/10.0+1.0e-4); dec2rgpulse(pwN,one,0.0,0.0); zgradpulse(gzlvl6, gt6); txphase(zero); delay(lambda-pwS6*0.5-gt6); h_sim3shapedpulse("reburp",shbw,shofs,0.0,2.0*pwN, zero, zero, zero, 0.0, 0.0); zgradpulse(gzlvl6, gt6); delay(lambda-pwS6*0.5-gt6); dec2rgpulse(pwN,t5,0.0,0.0); /**************************************************************************/ zgradpulse(-icosel*gzlvl2, gt1/10.0); dec2power(dpwr2); lk_sample(); if (n15_flg[0] =='y') { setreceiver(t14); } else { setreceiver(t24); } rcvron(); statusdelay(C,1.0e-4 ); }

pulsesequence() { // Define Variables and Objects and Get Parameter Values double pw1Xstmas = getval("pw1Xstmas"); double pw2Xstmas = getval("pw2Xstmas"); double tXzfselinit = getval("tXzfsel"); double tXzfsel = tXzfselinit - 3.0e-6; if (tXzfsel < 0.0) tXzfsel = 0.0; double d2init = getval("d2"); double d2 = d2init - pw1Xstmas/2.0 - pw2Xstmas/2.0; if (d2 < 0.0) d2 = 0.0; DSEQ dec = getdseq("H"); strncpy(dec.t.ch,"dec",3); putCmd("chHtppm='dec'\n"); strncpy(dec.s.ch,"dec",3); putCmd("chHspinal='dec'\n"); // Set Constant-time Period for d2. if (d2_index == 0) d2_init = getval("d2"); double d2_ = (ni - 1)/sw1 + d2_init; putCmd("d2acqret = %f\n",roundoff(d2_,12.5e-9)); putCmd("d2dwret = %f\n",roundoff(1.0/sw1,12.5e-9)); //-------------------------------------- // Copy Current Parameters to Processed //------------------------------------- putCmd("groupcopy('current','processed','acquisition')"); // Dutycycle Protection DUTY d = init_dutycycle(); d.dutyon = getval("pw1Xstmas") + getval("pw2Xstmas") + getval("pwXzfsel"); d.dutyoff = d1 + 4.0e-6; d.c1 = d.c1 + (!strcmp(dec.seq,"tppm")); d.c1 = d.c1 + ((!strcmp(dec.seq,"tppm")) && (dec.t.a > 0.0)); d.t1 = d2_ + tXzfsel + getval("rd") + getval("ad") + at; d.c2 = d.c2 + (!strcmp(dec.seq,"spinal")); d.c2 = d.c2 + ((!strcmp(dec.seq,"spinal")) && (dec.s.a > 0.0)); d.t2 = d2_ + tXzfsel + getval("rd") + getval("ad") + at; d = update_dutycycle(d); abort_dutycycle(d,10.0); // Set Phase Tables settable(ph1Xstmas,4,table1); settable(ph2Xstmas,4,table2); settable(phXzfsel,8,table3); settable(phRec,8,table4); if (phase1 == 2) { tsadd(ph1Xstmas,1,4); } setreceiver(phRec); // Begin Sequence txphase(ph1Xstmas); decphase(zero); obspower(getval("tpwr")); obspwrf(getval("aXstmas")); obsunblank(); decunblank(); _unblank34(); delay(d1); sp1on(); delay(2.0e-6); sp1off(); delay(2.0e-6); // H Decoupler on Before STMAS _dseqon(dec); // Two-Pulse STMAS rgpulse(getval("pw1Xstmas"),ph1Xstmas,0.0,0.0); txphase(ph2Xstmas); delay(d2); rgpulse(getval("pw2Xstmas"),ph2Xstmas,0.0,0.0); // Z-filter Pulse txphase(phXzfsel); obsblank(); obspower(getval("dbXzfsel")); obspwrf(getval("aXzfsel")); delay(3.0e-6); obsunblank(); delay(tXzfsel); rgpulse(getval("pwXzfsel"),phXzfsel,0.0,0.0); // Begin Acquisition obsblank(); _blank34(); delay(getval("rd")); startacq(getval("ad")); acquire(np, 1/sw); endacq(); _dseqoff(dec); obsunblank(); decunblank(); _unblank34(); }

void pulsesequence() { char codec[MAXSTR], codecseq[MAXSTR]; int icosel, t1_counter; double d2_init = 0.0, rf0 = 4095, rf200, pw200, copwr, codmf, cores, copwrf, tpwrs, pwHs = getval("pwHs"), compH = getval("compH"), pwClvl = getval("pwClvl"), pwC = getval("pwC"), compC = getval("compC"), pwNlvl = getval("pwNlvl"), pwN = getval("pwN"), sw1 = getval("sw1"), swH = getval("swH"), swC = getval("swC"), swN = getval("swN"), swTilt, angle_H = getval("angle_H"), angle_C = getval("angle_C"), angle_N, cos_H, cos_C, cos_N, mix = getval("mix"), tauCH = getval("tauCH"), /* 1/4JCH */ tauNH = getval("tauNH"), /* 1/4JNH */ tau1, tau2, tau3, tofali =getval("tofali"), /* aliphatic protons offset */ dofcaco =getval("dofcaco"), /* offset for caco decoupling, ~120 ppm */ dof = getval("dof"), gstab = getval("gstab"), gt0 = getval("gt0"), gzlvl0 = getval("gzlvl0"), gt1 = getval("gt1"), gzlvl1 = getval("gzlvl1"), gzlvl2 = getval("gzlvl2"), gt3 = getval("gt3"), gzlvl3 = getval("gzlvl3"), gt4 = getval("gt4"), gzlvl4 = getval("gzlvl4"), gt5 = getval("gt5"), gzlvl5 = getval("gzlvl5"), gt6 = getval("gt6"), gzlvl6 = getval("gzlvl6"), gt7 = getval("gt7"), gzlvl7 = getval("gzlvl7"), gt8 = getval("gt8"), gzlvl8 = getval("gzlvl8"), gt9 = getval("gt9"), gzlvl9 = getval("gzlvl9"); /* LOAD VARIABLES */ copwr = getval("copwr"); copwrf = getval("copwrf"); codmf = getval("codmf"); cores = getval("cores"); getstr("codecseq", codecseq); getstr("codec", codec); /* Load phase cycling variables */ settable(t1, 4, phi1); settable(t2, 2, phi2); settable(t3, 1, phi3); settable(t4, 8, phi4); settable(t5, 1, phi5); settable(t14, 2, phi14); settable(t11, 8, rec); angle_N=0.0; cos_N=0.0; /* activate auto-calibration flags */ setautocal(); if (autocal[0] == 'n') { /* 180 degree adiabatic C13 pulse from 0 to 200 ppm */ pw200 = getval("pw200"); rf200 = (compC*4095.0*pwC*4000.0*sqrt((30.0*sfrq/600.0+7.0)/0.35)); rf200 = (int) (rf200 + 0.5); if (1.0/(4000.0*sqrt((30.0*sfrq/600.0+7.0)/0.35)) < pwC ) { text_error( " Not enough C13 RF. pwC must be %f usec or less.\n", (1.0e6/(4000.0*sqrt((30.0*sfrq/600.0+7.0)/0.35))) ); psg_abort(1); } } else /* if autocal = 'y'(yes), 'q'(quiet), r(read), or 's'(semi) */ { strcpy(codecseq,"Pdec_154p"); if(FIRST_FID) /* call Pbox */ { ppm = getval("dfrq"); bw = 200.0*ppm; pws = 0.001; ofs = 0.0; nst = 1000; /* 1 ms long pulse, nst: number of steps */ stC200 = pbox_makeA("stC200", "sech", bw, pws, ofs, compC*pwC, pwClvl, nst); bw=20.0*ppm; ofs=154*ppm; Pdec_154p = pbox_Dsh("Pdec_154p", "WURST2", bw, ofs, compC*pwC, pwClvl); } rf200 = stC200.pwrf; pw200 = stC200.pw; } /* selective H20 one-lobe sinc pulse */ tpwrs = tpwr - 20.0*log10(pwHs/(compH*pw*1.69)); tpwrs = (int)(tpwrs+0.5); /* check validity of parameter range */ if((dm[A] == 'y' || dm[B] == 'y' || dm[C] == 'y')) { printf("incorrect Dec1 decoupler flags! "); psg_abort(1); } if((dm2[A] == 'y' || dm2[B] == 'y')) { printf("incorrect Dec2 decoupler flags! "); psg_abort(1); } if ((dpwr > 48) || (dpwr2 > 48)) { printf("don't fry the probe, dpwr too high! "); psg_abort(1); } /* set up angles for PR42 experiments */ /* sw1 is used as symbolic index */ if ( sw1 < 1000 ) { printf ("Please set sw1 to some value larger than 1000.\n"); psg_abort(1); } if (angle_H < 0 || angle_C < 0 || angle_H > 90 || angle_C > 90 ) { printf("angles must be set between 0 and 90 degree.\n"); psg_abort(1); } cos_H = cos (PI*angle_H/180); cos_C = cos (PI*angle_C/180); if ( (cos_H*cos_H + cos_C*cos_C) > 1.0) { printf ("Impossible angle combinations.\n"); psg_abort(1); } else { cos_N = sqrt(1 - (cos_H*cos_H + cos_C*cos_C) ); angle_N = acos(cos_N)*180/PI; } if (ix == 1) d2_init = d2; t1_counter = (int)((d2-d2_init)*sw1 + 0.5); if(t1_counter % 2) { tsadd(t3,2,4); tsadd(t11,2,4); } swTilt = swH*cos_H + swC*cos_C + swN*cos_N; if (phase1 == 1) {;} /* CC */ else if (phase1 == 2) { tsadd(t1, 1, 4); } /* SC */ else if (phase1 == 3) { tsadd(t2, 1, 4); tsadd(t14,1,4); } /* CS */ else if (phase1 == 4) { tsadd(t1, 1, 4); tsadd(t2,1,4); tsadd(t14,1,4); } /* SS */ if ( phase2 == 1 ) { tsadd(t5,2,4); icosel = 1; } else icosel = -1; tau1 = 1.0 * t1_counter * cos_H / swTilt; tau2 = 1.0 * t1_counter * cos_C / swTilt; tau3 = 1.0 * t1_counter * cos_N / swTilt; tau1 = tau1/2.0; tau2 = tau2/2.0; tau3 =tau3/2.0; if (ix ==1 ) { printf ("Current Spectral Width:\t\t%5.2f\n", swTilt); printf ("Angle_H: %5.2f \n", angle_H); printf ("Angle_C: %5.2f \n", angle_C); printf ("Angle_N: %5.2f \n", angle_N); printf ("\n\n\n\n\n"); } /* BEGIN ACTUAL PULSE SEQUENCE */ status(A); delay(d1); rcvroff(); obsoffset(tofali); obspower(tpwr); obspwrf(4095.0); decoffset(dof); decpower(pwClvl); decpwrf(rf0); dec2offset(dof2); dec2power(pwNlvl); dec2pwrf(4095.0); if (gt0 > 0.2e-6) { decrgpulse(pwC,zero,10.0e-6,0.0); dec2rgpulse(pwN,zero,2.0e-6,2.0e-6); zgradpulse(gzlvl0,gt0); delay(gstab); } txphase(t1); decphase(t2); dec2phase(zero); status(B); rgpulse(pw,t1,4.0e-6,2.0e-6); zgradpulse(gzlvl3,gt3); delay(2.0*tauCH - gt3 - 2.0*GRADIENT_DELAY -4.0e-6); decrgpulse(pwC,t2,2.0e-6,2.0e-6); decphase(zero); /*======= Start of c13 evolution ==========*/ if ( ((tau2 -PRG_START_DELAY - POWER_DELAY -pwN - 2.0*pwC/PI -2.0e-6)> 0) && ((tau2 -PRG_STOP_DELAY - POWER_DELAY - pwN - 2.0*pwC/PI -2.0e-6)>0) && (codec[A] == 'y') ) { decpower(copwr); decpwrf(copwrf); decprgon(codecseq,1/codmf,cores); decon(); delay(tau2 -PRG_START_DELAY - POWER_DELAY -pwN - 2.0*pwC/PI -2.0e-6); sim3pulse(2.0*pw, 0.0, 2.0*pwN, t1, zero, zero, 0.0, 0.0); delay(tau2 -PRG_STOP_DELAY - POWER_DELAY - pwN - 2.0*pwC/PI -2.0e-6); decoff(); decprgoff(); } else if ( (tau2 -pwN - 2.0*pwC/PI -2.0e-6) > 0) { delay(tau2 -pwN - 2.0*pwC/PI -2.0e-6); sim3pulse(2.0*pw, 0.0, 2.0*pwN, t1, zero, zero, 0.0, 0.0); delay(tau2 -pwN - 2.0*pwC/PI -2.0e-6); } else { delay(2.0*tau2); decphase(t14); delay(4.0e-6); sim3pulse(2.0*pw, 2.0*pwC, 2.0*pwN, t1, t14, zero, 0.0, 0.0); delay(4.0e-6); } decpower(pwClvl); decpwrf(rf0); decphase(zero); /*======= End of c13 evolution ==========*/ decrgpulse(pwC,zero, 2.0e-6,2.0e-6); txphase(zero); delay(2.0*tauCH + tau1 - gt3 - 4.0*pwC - gstab -4.0e-6 - 2.0*GRADIENT_DELAY); zgradpulse(gzlvl3,gt3); delay(gstab); decrgpulse(pwC,zero,0.0, 0.0); decphase(one); decrgpulse(2.0*pwC, one, 0.2e-6, 0.0); decphase(zero); decrgpulse(pwC, zero, 0.2e-6, 0.0); delay(tau1); rgpulse(pw,zero,2.0e-6,0.0); /* ===========Beginning of NOE transfer ======= */ delay(mix - gt4 - gt5 - pwN -pwC - 2.0*gstab); obsoffset(tof); zgradpulse(gzlvl4,gt4); delay(gstab); decrgpulse(pwC, zero, 2.0e-6, 2.0e-6); dec2rgpulse(pwN, zero, 2.0e-6, 2.0e-6); zgradpulse(gzlvl5,gt5); delay(gstab); /* H2O relaxes back to +z */ /* =========== End of NOE transfer ======= */ rgpulse(pw, zero, 2.0e-6, 2.0e-6); zgradpulse(gzlvl6,gt6); delay(tauNH - gt6 - 4.0e-6 - 2.0*GRADIENT_DELAY); sim3pulse(2.0*pw, 0.0, 2*pwN, zero, zero, zero, 2.0e-6, 2.0e-6); delay(tauNH - gt6 - gstab -4.0e-6 - 2.0*GRADIENT_DELAY); zgradpulse(gzlvl6,gt6); txphase(one); delay(gstab); rgpulse(pw,one,2.0e-6,2.0e-6); txphase(two); obspower(tpwrs); shaped_pulse("H2Osinc", pwHs, two, 2.0e-6, 2.0e-6); obspower(tpwr); zgradpulse(gzlvl7,gt7); dec2phase(t3); decoffset(dofcaco); /* offset on 120ppm for CaCO decoupling */ decpwrf(rf200); /* fine power for stC200 */ delay(gstab); dec2rgpulse(pwN,t3,2.0e-6,2.0e-6); dec2phase(t4); delay(tau3); rgpulse(2.0*pw, zero, 2.0e-6, 2.0e-6); decshaped_pulse("stC200", 1.0e-3, zero, 2.0e-6, 2.0e-6); delay(tau3); delay(gt1 +gstab +8.0e-6 - 1.0e-3 - 2.0*pw); dec2rgpulse(2.0*pwN, t4, 2.0e-6, 2.0e-6); dec2phase(t5); zgradpulse(gzlvl1, gt1); delay(gstab + WFG_START_DELAY + WFG_STOP_DELAY - 2.0*GRADIENT_DELAY); sim3pulse(pw, 0.0, pwN, zero, zero, t5, 2.0e-6, 2.0e-6); dec2phase(zero); zgradpulse(gzlvl8, gt8); delay(tauNH - gt8 - 2.0*GRADIENT_DELAY -4.0e-6); sim3pulse(2.0*pw, 0.0, 2.0*pwN, zero, zero, zero, 2.0e-6, 2.0e-6); delay(tauNH - gt8 - gstab -4.0e-6 - 2.0*GRADIENT_DELAY); zgradpulse(gzlvl8, gt8); txphase(one); dec2phase(one); delay(gstab); sim3pulse(pw, 0.0, pwN, one, zero, one, 2.0e-6, 2.0e-6); txphase(zero); dec2phase(zero); zgradpulse(gzlvl9, gt9); delay(tauNH - gt9 - 2.0*GRADIENT_DELAY -4.0e-6); sim3pulse(2.0*pw, 0.0, 2.0*pwN, zero, zero, zero, 2.0e-6, 2.0e-6); delay(tauNH - gt9 - 2.0*GRADIENT_DELAY -gstab -4.0e-6); zgradpulse(gzlvl9, gt9); delay(gstab); rgpulse(pw, zero, 2.0e-6, 2.0e-6); delay((gt1/10.0) +gstab -2.0e-6 - 0.5*pw + 2.0*GRADIENT_DELAY + POWER_DELAY); rgpulse(2.0*pw, zero, 2.0e-6, 2.0e-6); zgradpulse(icosel*gzlvl2, gt1/10.0); dec2power(dpwr2); delay(gstab); status(C); setreceiver(t11); }

pulsesequence() { /* DECLARE AND LOAD VARIABLES */ char CA90_in_str[MAXSTR], CA180_in_str[MAXSTR], CA180n_in_str[MAXSTR], CO180offCA_in_str[MAXSTR], RFpars[MAXSTR], exp_mode[MAXSTR], /* flag to run 3D, or 2D time-shared 15N TROSY /13C HSQC-SE*/ f1180[MAXSTR], /* Flag to start t1 @ halfdwell */ f2180[MAXSTR], f3180[MAXSTR]; /* do TROSY on N15 and H1 */ int icosel=1.0; /* used to get n and p type */ double x,y,z, t2max, t1max, tpwrs, tpwrsf_d = getval("tpwrsf_d"), /* fine power adustment for first soft pulse(down)*/ tpwrsf_u = getval("tpwrsf_u"), /* fine power adustment for second soft pulse(up) */ pwHs = getval("pwHs"), /* H1 90 degree pulse length at tpwrs */ compH =getval("compH"), tau1, tau2, /*evolution times in indirect dimensions */ ni2=getval("ni2"), tauNH=getval("tauNH"), /* 1/(4Jhn), INEPTs, 2.4ms*/ tauNH1=getval("tauNH1"), /* 1/(4Jhn), TROSY in CN CT, 2.7ms*/ timeTN1=getval("timeTN1"), /* CT time for (first) N->CA*N transfer */ timeTN=getval("timeTN"), /* CT time for last SE TROSY */ timeCN=getval("timeCN"), /* CT time for CA -> N transfer, middle */ pwClvl = getval("pwClvl"), /* coarse power for C13 pulse */ pwC = getval("pwC"), /* C13 90 degree pulse length at pwClvl */ compC = getval("compC"), dfrq = getval("dfrq"), pwNlvl = getval("pwNlvl"), /* power for N15 pulses */ pwN = getval("pwN"), /* N15 90 degree pulse length at pwNlvl */ gstab = getval("gstab"), g6bal= getval("g6bal"), /* balance of the decoding gradient around last 180 pulse on 1H g6bal=1.0 : full g6 is on the right side of the last pw180 on 1H g6bal=0.0: full g6 is on the left side*/ gt0 = getval("gt0"), gt1 = getval("gt1"), gt2 = getval("gt2"), gt3 = getval("gt3"), gt4 = getval("gt4"), gt5 = getval("gt5"), gt6 = getval("gt6"), gt7 = getval("gt7"), gzlvl0 = getval("gzlvl0"), gzlvl1 = getval("gzlvl1"), gzlvl2 = getval("gzlvl2"), gzlvl3 = getval("gzlvl3"), gzlvl4 = getval("gzlvl4"), gzlvl5 = getval("gzlvl5"), gzlvl6 = getval("gzlvl6"), gzlvl7 = getval("gzlvl7"), gzlvl11 = getval("gzlvl11"); getstr("f1180",f1180); getstr("f2180",f2180); getstr("exp_mode",exp_mode); tpwrs = tpwr - 20.0*log10(pwHs/(compH*pw*1.69)); /*needs 1.69 times more*/ tpwrs = (int) (tpwrs); /*power than a square pulse */ if (tpwrsf_d<4095.0) tpwrs=tpwrs+6.0; /* add 6dB to let tpwrsf_d control fine power ~2048*/ /* LOAD PHASE TABLE */ settable(t1,1,phi1); settable(t2,4,phi2); /* default double trosy */ if (exp_mode[A] == 'h') {settable(t2,4,phi2h);}; /*option for regular hNcaNH */ settable(t3,4,phi3); settable(t4,8,phi4); settable(t5,2,phi5); settable(t6,4,phi6); settable(t7,4,phi7); settable(t8,4,phi8); settable(t21,1,psi1); /*trosy and SE hsqc in reverse INPET */ settable(t22,1,psi2); settable(t23,1,psi2c); settable(t31,8,rec); /* some checks */ if((dm2[A] == 'y') || (dm2[B] == 'y') || (dm2[C] == 'y') || (dm2[D] == 'y')) { text_error("incorrect dec2 decoupler flags! Should be 'nnnn' "); psg_abort(1); } if ( dm3[A] == 'y' || dm3[C] == 'y' ) { printf("incorrect dec3 decoupler flags! Should be 'nyn' or 'nnn' "); psg_abort(1);} if ( dpwr3 > 56 ) { printf("dpwr3 too large! recheck value "); psg_abort(1);} if ( (dm3[B] == 'y' ) && (timeCN*2.0 > 60.0e-3) ) { printf("too lond time for 2H decoupling, SOL ");psg_abort(1);} /* INITIALIZE VARIABLES */ if(FIRST_FID) /* call Pbox */ { getstr("CA180_in_str",CA180_in_str); getstr("CA180n_in_str",CA180n_in_str); getstr("CA90_in_str",CA90_in_str); getstr("CO180offCA_in_str",CO180offCA_in_str); strcpy(RFpars, "-stepsize 0.5 -attn i"); CA180 = pbox("et_CA180_auto", CA180_in_str, RFpars, dfrq, compC*pwC, pwClvl); CA180n = pbox("et_CA180n_auto", CA180n_in_str, RFpars, dfrq, compC*pwC, pwClvl); CA90 = pbox("et_CA90_auto", CA90_in_str, RFpars, dfrq, compC*pwC, pwClvl); CO180offCA = pbox("et_CO180offCA_auto", CO180offCA_in_str, RFpars, dfrq, compC*pwC, pwClvl); }; /* Phase incrementation for hypercomplex 2D data, States-Haberkorn element */ /* t1 , N15 */ if (phase1 == 2) {tsadd(t2 ,1,4);} if(d2_index % 2) {tsadd(t2,2,4); tsadd(t31,2,4); } /* setting up semi-CT on t1 (ni) dimension */ tau1 = d2; t1max=(ni-1.0)/sw1; if((f1180[A] == 'y') && (ni > 0.0)) {tau1 += 0.5/sw1 ; t1max+= 0.5/sw1; } if( t1max < timeTN1*2.0) {t1max=2.0*timeTN1;}; /* if not enough ni increments, then just regular CT in t1/ni CN */ /* t2, CA */ if (phase2 == 2) { tsadd(t3,1,4); } if (d3_index % 2) { tsadd(t3,2,4); tsadd(t31,2,4); } /* setup constant time in t2 (ni2) */ tau2 = d3; t2max=2.0*(timeCN - CO180offCA.pw); if((f2180[A] == 'y') && (ni2 > 0.0)) {tau2 += 0.5/sw2 ; t2max += 0.5/sw2 ;} if(tau2 < 0.2e-6) {tau2 = 0.0;} if ( (ni2-1.0)/sw2 > t2max) { text_error("too many ni2 increments in t2 ! "); psg_abort(1); } if(FIRST_FID) { printf("t1max is %f\n",t1max); printf("t2max is %f\n",t2max); }; /* BEGIN PULSE SEQUENCE */ status(A); obspower(tpwr); decpower(pwClvl); dec2power(pwNlvl); txphase(zero); decphase(zero); dec2phase(zero); delay(d1); zgradpulse(gzlvl2, gt2); delay(gstab*3.0); if (exp_mode[B]=='n') dec2rgpulse(2.0*pwN, zero, 0.0, 0.0); /* test for steady-state 15N */ /* Hz -> HzXz INEPT */ rgpulse(pw,zero,rof1,rof1); /* 1H pulse excitation */ zgradpulse(gzlvl0, gt0); delay(tauNH -gt0); sim3pulse(2.0*pw, 0.0, 2.0*pwN, zero, zero, zero, 0.0, 0.0); delay(tauNH - gt0 -gstab); zgradpulse(gzlvl0, gt0); delay(gstab); rgpulse(pw, t6, rof1, rof1); /* on HzNz now */ /* water flipback*/ obspower(tpwrs); obspwrf(tpwrsf_u); shaped_pulse("H2Osinc",pwHs,zero,rof1,rof1); obspower(tpwr); obspwrf(4095.0); /* purge */ zgradpulse(gzlvl3, gt3); dec2phase(t2); delay(gstab*2.0); /* HzNz -> NzCAz +t1 evolution*/ dec2rgpulse(pwN, t2, 0.0, 0.0); /* double-trosy hNcaNH */ delay(tauNH1 -pwHs-4.0*rof1 -pw -2.0*POWER_DELAY -WFG_STOP_DELAY-WFG_START_DELAY); obspower(tpwrs); obspwrf(tpwrsf_d); shaped_pulse("H2Osinc",pwHs,two,rof1,rof1); obspower(tpwr); obspwrf(4095.0); rgpulse(pw, zero, rof1, rof1); rgpulse(pw, t7, rof1, rof1); obspower(tpwrs); obspwrf(tpwrsf_u); shaped_pulse("H2Osinc",pwHs,t8,rof1,rof1); obspower(tpwr); obspwrf(4095.0); dec_c13_shpulse(CO180offCA,zero); delay(tau1*0.5); dec_c13_shpulse(CO180offCA,zero); dec2phase(zero); delay( timeTN1 -tauNH1 -pwHs -4.0*rof1 -pw -2.0*POWER_DELAY -WFG_STOP_DELAY -WFG_START_DELAY -CA180.pw -2.0*CO180offCA.pw -3.0*(2.0*POWER_DELAY +WFG_STOP_DELAY +WFG_START_DELAY)); dec_c13_shpulse(CA180,zero); delay(tau1*0.5 -timeTN1*tau1/t1max); dec2rgpulse(2.0*pwN, zero, 0.0, 0.0); delay( timeTN1 -tau1*timeTN1/t1max); dec2rgpulse(pwN, zero, 0.0, 0.0); /* on CAzNz now */ /* purge */ zgradpulse(gzlvl7, gt7); delay(gstab); if(dm3[B] == 'y') { dec3unblank(); if(1.0/dmf3>900.0e-6) { dec3power(dpwr3+6.0); dec3rgpulse(0.5/dmf3, one, 1.0e-6, 0.0e-6); dec3power(dpwr3); } else dec3rgpulse(1.0/dmf3, one, 1.0e-6,0.0e-6); dec3phase(zero); setstatus(DEC3ch, TRUE, 'w', FALSE, dmf3); } /* dec_c13_shpulse(CA90,t3);*/ /* t2 time, CA evolution */ decrgpulse(pwC,t3,0.0,0.0); decphase(zero); delay(0.5*(timeCN+tau2*0.5-CO180offCA.pw) ); dec_c13_shpulse(CO180offCA,zero); delay(0.5*(timeCN+tau2*0.5-CO180offCA.pw) -pwN*2.0 + WFG_STOP_DELAY); if (exp_mode[A]=='R') /* test CA.N relaxation rate */ { delay(2.0*pwN); } else dec2rgpulse(2.0*pwN, zero, 0.0, 0.0); dec_c13_shpulse(CA180n,zero); delay(0.5*(timeCN-tau2*0.5-CO180offCA.pw) ); dec_c13_shpulse(CO180offCA,zero); delay(0.5*(timeCN-tau2*0.5-CO180offCA.pw) + WFG_START_DELAY); /*dec_c13_shpulse(CA90,zero);*/ decrgpulse(pwC,zero,0.0,0.0); if(dm3[B] == 'y') { setstatus(DEC3ch, FALSE, 'w', FALSE, dmf3); if(1.0/dmf3>900.0e-6) { dec3power(dpwr3+6.0); dec3rgpulse(0.5/dmf3, three, 1.0e-6, 0.0e-6); dec3power(dpwr3); } else dec3rgpulse(1.0/dmf3, three, 1.0e-6, 0.0e-6); dec3blank(); delay(PRG_START_DELAY); } zgradpulse(gzlvl5, gt5); dec2phase(t4); delay(gstab); /* CaN->N + back to NH */ dec2rgpulse(pwN, t4, 0.0, 0.0); dec2phase(zero); delay(timeTN); dec2rgpulse(2.0*pwN, zero, 0.0, 0.0); dec_c13_shpulse(CA180,zero); delay(timeTN - CA180.pw -gt4-gstab -pwHs-3.0*rof1 -4.0*POWER_DELAY -2.0*WFG_STOP_DELAY-2.0*WFG_START_DELAY -2.0*GRADIENT_DELAY +4.0*pwN/3.1415-pw); zgradpulse(gzlvl4, gt4); delay(gstab); /*Water flipback (flipdown actually ) */ obspower(tpwrs); obspwrf(tpwrsf_d); shaped_pulse("H2Osinc",pwHs,three,rof1,rof1); obspower(tpwr); obspwrf(4095.0); /* reverse double INEPT */ /* 90 */ rgpulse(pw, t21, rof1, rof1); zgradpulse(gzlvl11, gt1); delay(tauNH -gt1 -rof1 -CA180.pw -2.0*POWER_DELAY - WFG_STOP_DELAY- WFG_START_DELAY ); dec_c13_shpulse(CA180,zero); sim3pulse(2.0*pw, 0.0, 2.0*pwN, zero, zero, zero, 0.0, 0.0); delay(tauNH - gt1 -gstab); zgradpulse(gzlvl11, gt1); delay(gstab); /* 90 */ sim3pulse(pw, 0.0, pwN, one, zero, zero, 0.0, 0.0); zgradpulse(gzlvl1, gt1); delay(tauNH -gt1); sim3pulse(2.0*pw, 0.0, 2.0*pwN, zero, zero, zero, 0.0, 0.0); delay(tauNH -POWER_DELAY -gt1- gstab); zgradpulse(gzlvl1, gt1); dec2phase(t22); delay(gstab); sim3pulse(0.0,0.0, pwN, one, zero, t22, 0.0, 0.0); zgradpulse(-(1.0-g6bal)*gzlvl6*icosel, gt6); /* 2.0*GRADIENT_DELAY */ delay( gstab -pwN*0.5 +pw*(2.0/3.1415-0.5) ); rgpulse(2.0*pw, zero, rof1, rof1); dec2power(dpwr2); decpower(dpwr); zgradpulse(g6bal*gzlvl6*icosel, gt6); /* 2.0*GRADIENT_DELAY */ delay(gstab +2.0*POWER_DELAY ); status(C); setreceiver(t31); }

pulsesequence() { /* DECLARE AND LOAD VARIABLES */ char f1180[MAXSTR], /* Flag to start t1 @ halfdwell */ f2180[MAXSTR], /* Flag to start t2 @ halfdwell */ mag_flg[MAXSTR], /* magic-angle coherence transfer gradients */ SCT[MAXSTR], /* Semi-constant time flag for N-15 evolution */ CT_c[MAXSTR], /* Constant time flag for C-13 evolution */ TROSY[MAXSTR]; /* do TROSY on N15 and H1 */ int icosel, /* used to get n and p type */ t1_counter, /* used for states tppi in t1 */ t2_counter, /* used for states tppi in t2 */ PRexp, /* projection-reconstruction flag */ ni2 = getval("ni2"); double tau1, /* t1 delay */ tau2, /* t2 delay */ tauC = getval("tauC"), /* delay for CO to Ca evolution */ timeTN = getval("timeTN"), /* constant time for 15N evolution */ timeTC = getval("timeTC"), /* constant time for 13C evolution */ t2a=0.0, t2b=0.0, halfT2=0.0, CTdelay=0.0, kappa = 5.4e-3, lambda = 2.4e-3, pwClvl = getval("pwClvl"), /* coarse power for C13 pulse */ pwC = getval("pwC"), /* C13 90 degree pulse length at pwClvl */ rf0, /* maximum fine power when using pwC pulses */ /* 90 degree pulse at Ca (56ppm), first off-resonance null at CO (174ppm) */ pwC1, /* 90 degree pulse length on C13 at rf1 */ rf1, /* fine power for 4.7 kHz rf for 600MHz magnet */ /* 180 degree pulse at Ca (56ppm), first off-resonance null at CO(174ppm) */ pwC2, /* 180 degree pulse length at rf2 */ rf2, /* fine power for 10.5 kHz rf for 600MHz magnet */ /* the following pulse lengths for SLP pulses are automatically calculated */ /* by the macro "proteincal". SLP pulse shapes, "offC6" etc are called */ /* directly from your shapelib. */ pwC3 = getval("pwC3"), /*180 degree pulse at Ca(56ppm) null at CO(174ppm) */ pwC6 = getval("pwC6"), /* 90 degree selective sinc pulse on CO(174ppm) */ pwC8 = getval("pwC8"), /* 180 degree selective sinc pulse on CO(174ppm) */ pwC9 = getval("pwC9"), /* 180 degree selective sinc pulse on CO(174ppm) */ phshift9, /* phase shift induced on Ca by pwC9 ("offC9") pulse */ pwZ, /* the largest of pwC9 and 2.0*pwN */ pwZ1, /* the larger of pwC8 and 2.0*pwN for 1D experiments */ rf3, /* fine power for the pwC3 ("offC3") pulse */ rf6, /* fine power for the pwC6 ("offC6") pulse */ rf8, /* fine power for the pwC8 ("offC8") pulse */ rf9, /* fine power for the pwC9 ("offC9") pulse */ dofCO, /* channel 2 offset for most CO pulses */ compH = getval("compH"), /* adjustment for C13 amplifier compression */ compC = getval("compC"), /* adjustment for C13 amplifier compression */ pwHs = getval("pwHs"), /* H1 90 degree pulse length at tpwrs */ tpwrsf = getval("tpwrsf"), /* fine power adjustment for flipback */ tpwrs, /* power for the pwHs ("H2Osinc") pulse */ csa, sna, pra = M_PI*getval("pra")/180.0, pwHd, /* H1 90 degree pulse length at tpwrd */ tpwrd, /* rf for WALTZ decoupling */ waltzB1 = getval("waltzB1"), /* waltz16 field strength (in Hz) */ pwNlvl = getval("pwNlvl"), /* power for N15 pulses */ pwN = getval("pwN"), /* N15 90 degree pulse length at pwNlvl */ sw1 = getval("sw1"), sw2 = getval("sw2"), gt1 = getval("gt1"), /* coherence pathway gradients */ gzcal = getval("gzcal"), /* g/cm to DAC conversion factor */ gzlvl1 = getval("gzlvl1"), gzlvl2 = getval("gzlvl2"), gt0 = getval("gt0"), /* other gradients */ gt3 = getval("gt3"), gt4 = getval("gt4"), gt5 = getval("gt5"), gt7 = getval("gt7"), gt9 = getval("gt9"), gt10 = getval("gt10"), gstab = getval("gstab"), gzlvl0 = getval("gzlvl0"), gzlvl3 = getval("gzlvl3"), gzlvl4 = getval("gzlvl4"), gzlvl5 = getval("gzlvl5"), gzlvl6 = getval("gzlvl6"), gzlvl7 = getval("gzlvl7"), gzlvl8 = getval("gzlvl8"), gzlvl9 = getval("gzlvl9"), gzlvl10 = getval("gzlvl10"); getstr("f1180",f1180); getstr("f2180",f2180); getstr("mag_flg",mag_flg); getstr("SCT",SCT); getstr("CT_c",CT_c); getstr("TROSY",TROSY); /* LOAD PHASE TABLE */ settable(t3,2,phi3); settable(t4,1,phx); settable(t5,4,phi5); if (TROSY[A]=='y') {settable(t8,1,phy); settable(t9,1,phx); settable(t10,1,phy); settable(t11,1,phx); settable(t12,4,recT);} else {settable(t8,1,phx); settable(t9,8,phi9); settable(t10,1,phx); settable(t11,1,phy); settable(t12,4,rec);} /* INITIALIZE VARIABLES */ if( dpwrf < 4095 ) { printf("reset dpwrf=4095 and recalibrate C13 90 degree pulse"); psg_abort(1); } /* offset during CO pulses, except for t1 evolution period */ dofCO = dof + 118.0*dfrq; /* maximum fine power for pwC pulses */ rf0 = 4095.0; /* 90 degree pulse on Ca, null at CO 118ppm away */ pwC1 = sqrt(15.0)/(4.0*118.0*dfrq); rf1 = 4095.0*(compC*pwC)/pwC1; rf1 = (int) (rf1 + 0.5); /* 180 degree pulse on Ca, null at CO 118ppm away */ pwC2 = sqrt(3.0)/(2.0*118.0*dfrq); rf2 = (compC*4095.0*pwC*2.0)/pwC2; rf2 = (int) (rf2 + 0.5); if( rf2 > 4095.0 ) { printf("increase pwClvl so that C13 90 < 24us*(600/sfrq)"); psg_abort(1);} /* 180 degree pulse on Ca, null at CO 118ppm away */ rf3 = (compC*4095.0*pwC*2.0)/pwC3; rf3 = (int) (rf3 + 0.5); /* 90 degree one-lobe sinc pulse on CO, null at Ca 118ppm away */ rf6 = (compC*4095.0*pwC*1.69)/pwC6; /* needs 1.69 times more */ rf6 = (int) (rf6 + 0.5); /* power than a square pulse */ /* 180 degree one-lobe sinc pulse on CO, null at Ca 118ppm away */ rf8 = (compC*4095.0*pwC*2.0*1.65)/pwC8; /* needs 1.65 times more */ rf8 = (int) (rf8 + 0.5); /* power than a square pulse */ /* 180 degree one-lobe sinc pulse on CO, null at Ca 118ppm away */ rf9 = (compC*4095.0*pwC*2.0*1.65)/pwC8; /* needs 1.65 times more */ rf9 = (int) (rf9 + 0.5); /* power than a square pulse */ /* the pwC9 pulse at the middle of t1 */ if ((ni2 > 0.0) && (ni == 1.0)) ni = 0.0; if (pwC8 > 2.0*pwN) pwZ = pwC8; else pwZ = 2.0*pwN; if ((pwC9==0.0) && (pwC8>2.0*pwN)) pwZ1=pwC8-2.0*pwN; else pwZ1=0.0; if ( ni > 1 ) pwC9 = pwC8; if ( pwC9 > 0 ) phshift9 = 140.0; else phshift9 = 0.0; /* selective H20 one-lobe sinc pulse */ tpwrs = tpwr - 20.0*log10(pwHs/(compH*pw*1.69)); /* needs 1.69 times more */ tpwrs = (int) (tpwrs); /* power than a square pulse */ /* power level and pulse time for WALTZ 1H decoupling */ pwHd = 1/(4.0 * waltzB1) ; /* 7.5 kHz rf */ tpwrd = tpwr - 20.0*log10(pwHd/(compH*pw)); tpwrd = (int) (tpwrd + 0.5); /* set up Projection-Reconstruction experiment */ tau1 = d2; tau2 = d3; PRexp=0; csa = 1.0; sna = 0.0; if((pra > 0.0) && (pra < 90.0)) /* PR experiments */ { PRexp = 1; csa = cos(pra); sna = sin(pra); tau1 = d2*csa; tau2 = d2*sna; } /* CHECK VALIDITY OF PARAMETER RANGES */ if(SCT[A] == 'n') { if (PRexp) { if( 0.5*ni*sna/sw1 > timeTN - WFG3_START_DELAY) { printf(" ni is too big. Make ni equal to %d or less.\n", ((int)((timeTN - WFG3_START_DELAY)*2.0*sw1/sna))); psg_abort(1);} } else { if ( 0.5*ni2*1/(sw2) > timeTN - WFG3_START_DELAY) { printf(" ni2 is too big. Make ni2 equal to %d or less.\n", ((int)((timeTN - WFG3_START_DELAY)*2.0*sw2))); psg_abort(1);} } } if(CT_c[A] == 'y') { if ( 0.5*ni*csa/sw1 > timeTC) { printf(" ni is too big. Make ni less than %d or less.\n", ((int)(timeTC*2.0*sw1/csa - 4e-6 - SAPS_DELAY))); psg_abort(1);} } if ( tauC < (gt7+1.0e-4+0.5*10.933*pwC)) gt7=(tauC-1.0e-4-0.5*10.933*pwC); if ( dm[A] == 'y' || dm[B] == 'y' || dm[C] == 'y' ) { printf("incorrect dec1 decoupler flags! Should be 'nnn' "); psg_abort(1);} if ( dm2[A] == 'y' || dm2[B] == 'y' ) { printf("incorrect dec2 decoupler flags! Should be 'nny' "); psg_abort(1);} if ( dm3[A] == 'y' || dm3[C] == 'y' ) { printf("incorrect dec3 decoupler flags! Should be 'nyn' or 'nnn' "); psg_abort(1);} if ( dpwr2 > 50 ) { printf("dpwr2 too large! recheck value "); psg_abort(1);} if ( pw > 20.0e-6 ) { printf(" pw too long ! recheck value "); psg_abort(1);} if ( pwN > 100.0e-6 ) { printf(" pwN too long! recheck value "); psg_abort(1);} if ( TROSY[A]=='y' && dm2[C] == 'y') { text_error("Choose either TROSY='n' or dm2='n' ! "); psg_abort(1);} /* PHASES AND INCREMENTED TIMES */ /* Phase incrementation for hypercomplex 2D data, States-Haberkorn element */ if (phase1 == 2) tsadd(t3,1,4); if (TROSY[A]=='y') { if (phase2 == 2) icosel = +1; else {tsadd(t4,2,4); tsadd(t10,2,4); icosel = -1;} } else { if (phase2 == 2) {tsadd(t10,2,4); icosel = +1;} else icosel = -1; } /* Calculate modifications to phases for States-TPPI acquisition */ if( ix == 1) d2_init = d2; t1_counter = (int) ( (d2-d2_init)*sw1 + 0.5 ); if(t1_counter % 2) { tsadd(t3,2,4); tsadd(t12,2,4); } if( ix == 1) d3_init = d3; t2_counter = (int) ( (d3-d3_init)*sw2 + 0.5 ); if(t2_counter % 2) { tsadd(t8,2,4); tsadd(t12,2,4); } /* Set up CONSTANT/SEMI-CONSTANT time evolution in N15 */ halfT2 = 0.0; CTdelay = timeTN + pwC8 + WFG_START_DELAY - SAPS_DELAY; if(ni>1) { if(f1180[A] == 'y') /* Set up f1180 */ tau1 += 0.5*csa/sw1; /* if not PRexp then csa = 1.0 */ if(PRexp) { halfT2 = 0.5*(ni-1)/sw1; /* ni2 is not defined */ if(f1180[A] == 'y') { tau2 += 0.5*sna/sw1; halfT2 += 0.25*sna/sw1; } t2b = (double) t1_counter*((halfT2 - CTdelay)/((double)(ni-1))); } } if (ni2>1) { halfT2 = 0.5*(ni2-1)/sw2; if(f2180[A] == 'y') /* Set up f2180 */ { tau2 += 0.5/sw2; halfT2 += 0.25/sw2; } t2b = (double) t2_counter*((halfT2 - CTdelay)/((double)(ni2-1))); } tau1 = tau1/2.0; tau2 = tau2/2.0; if(tau1 < 0.2e-6) tau1 = 0.0; if(tau2 < 0.2e-6) tau2 = 0.0; if(t2b < 0.0) t2b = 0.0; t2a = CTdelay - tau2 + t2b; if(t2a < 0.2e-6) t2a = 0.0; /* uncomment these lines to check t2a and t2b printf("%d: t2a = %.12f", t2_counter,t2a); printf(" ; t2b = %.12f\n", t2b); */ /* BEGIN PULSE SEQUENCE */ status(A); delay(d1); if ( dm3[B] == 'y' ) { lk_hold(); lk_sampling_off();} /*freezes z0 correction, stops lock pulsing*/ rcvroff(); obspower(tpwr); decpower(pwClvl); dec2power(pwNlvl); decpwrf(rf0); obsoffset(tof); decoffset(dofCO); txphase(zero); delay(1.0e-5); if (TROSY[A] == 'n') dec2rgpulse(pwN, zero, 0.0, 0.0); /*destroy N15 and C13 magnetization*/ decrgpulse(pwC, zero, 0.0, 0.0); zgradpulse(-gzlvl0, 0.5e-3); delay(1.0e-4); if (TROSY[A] == 'n') dec2rgpulse(pwN, one, 0.0, 0.0); decrgpulse(pwC, zero, 0.0, 0.0); zgradpulse(-0.7*gzlvl0, 0.5e-3); delay(5.0e-4); rgpulse(pw, zero, 0.0, 0.0); /* 1H pulse excitation */ dec2phase(zero); zgradpulse(gzlvl0, gt0); delay(lambda - gt0); sim3pulse(2.0*pw, 0.0, 2.0*pwN, zero, zero, zero, 0.0, 0.0); txphase(one); zgradpulse(gzlvl0, gt0); delay(lambda - gt0); rgpulse(pw, one, 0.0, 0.0); if (tpwrsf < 4095.0) {obspwrf(tpwrsf); obspower(tpwrs+6.0);} /* increases tpwrs by 6dB, now need */ else obspower(tpwrs); /* tpwrsf to be ~ 2048 for equivalence */ if (TROSY[A]=='y') {txphase(two); shaped_pulse("H2Osinc", pwHs, two, 5.0e-4, 0.0); obspower(tpwr); obspwrf(4095.0); zgradpulse(gzlvl3, gt3); delay(2.0e-4); dec2rgpulse(pwN, zero, 0.0, 0.0); delay(0.5*kappa - 2.0*pw); rgpulse(2.0*pw, two, 0.0, 0.0); obspower(tpwrd); /* POWER_DELAY */ decphase(zero); dec2phase(zero); decpwrf(rf8); delay(timeTN -0.5*kappa - POWER_DELAY - WFG3_START_DELAY); } else {txphase(zero); if (tpwrsf < 4095.0) {obspwrf(tpwrsf); obspower(tpwrs+6.0);} /* increases tpwrs by 6dB, now need */ else obspower(tpwrs); /* tpwrsf to be ~ 2048 for equivalence */ shaped_pulse("H2Osinc",pwHs,zero,5.0e-4,0.0); obspower(tpwrd); obspwrf(4095.0); zgradpulse(gzlvl3, gt3); delay(2.0e-4); dec2rgpulse(pwN, zero, 0.0, 0.0); txphase(one); delay(kappa - pwHd - 2.0e-6 - PRG_START_DELAY); rgpulse(pwHd,one,0.0,0.0); txphase(zero); delay(2.0e-6); obsprgon("waltz16", pwHd, 90.0); /* PRG_START_DELAY */ xmtron(); decphase(zero); dec2phase(zero); decpwrf(rf8); delay(timeTN - kappa - WFG3_START_DELAY); } /* WFG3_START_DELAY */ sim3shaped_pulse("", "offC8", "", 0.0, pwC8, 2.0*pwN, zero, zero, zero, 0.0, 0.0); decphase(zero); decpwrf(rf6); delay(timeTN); dec2rgpulse(pwN, zero, 0.0, 0.0); if (TROSY[A]=='n') {xmtroff(); obsprgoff(); rgpulse(pwHd,three,2.0e-6,0.0);} zgradpulse(-gzlvl3, gt3); delay(2.0e-4); decshaped_pulse("offC6", pwC6, zero, 0.0, 0.0); zgradpulse(-gzlvl7, gt7); decpwrf(rf0); decphase(zero); delay(tauC - gt7 - 0.5*10.933*pwC); decrgpulse(pwC*158.0/90.0, zero, 0.0, 0.0); decrgpulse(pwC*171.2/90.0, two, 0.0, 0.0); decrgpulse(pwC*342.8/90.0, zero, 0.0, 0.0); /* Shaka 6 composite */ decrgpulse(pwC*145.5/90.0, two, 0.0, 0.0); decrgpulse(pwC*81.2/90.0, zero, 0.0, 0.0); decrgpulse(pwC*85.3/90.0, two, 0.0, 0.0); zgradpulse(-gzlvl7, gt7); decpwrf(rf6); decphase(one); txphase(one); delay(tauC - gt7 - 0.5*10.933*pwC - WFG_START_DELAY); /* WFG_START_DELAY */ decshaped_pulse("offC6", pwC6, one, 0.0, 0.0); decoffset(dof); zgradpulse(-gzlvl9, gt9); decpwrf(rf1); decphase(t3); delay(2.0e-4); if ( dm3[B] == 'y' ) /* begins optional 2H decoupling */ { dec3rgpulse(1/dmf3,one,10.0e-6,2.0e-6); dec3unblank(); dec3phase(zero); delay(2.0e-6); setstatus(DEC3ch, TRUE, 'w', FALSE, dmf3); } rgpulse(pwHd,one,0.0,0.0); txphase(zero); delay(2.0e-6); obsprgon("waltz16", pwHd, 90.0); xmtron(); decrgpulse(pwC1, t3, 0.0, 0.0); decphase(zero); /* xxxxxxxxxxxxxxxxxxxxxx 13Ca EVOLUTION xxxxxxxxxxxxxxxxxx */ if (ni==1.0) /* special 1D check of pwC9 phase enabled when ni=1 */ { decpwrf(rf9); delay(10.0e-6 + SAPS_DELAY + 0.5*pwZ1); /* WFG3_START_DELAY */ sim3shaped_pulse("", "offC9", "", 0.0, pwC9, 2.0*pwN, zero, zero, zero, 2.0e-6, 0.0); initval(phshift9, v9); decstepsize(1.0); dcplrphase(v9); /* SAPS_DELAY */ delay(10.0e-6 + WFG3_START_DELAY + 0.5*pwZ1); } else if(CT_c[A] == 'y') /* xxxxxxx 13Ca Constant Time EVOLUTION xxxxxxxx */ { decpwrf(rf9); if(tau1 - 2.0*pwC1/PI - WFG_START_DELAY -POWER_DELAY > 0.0) { delay(tau1 -2.0*pwC1/PI -POWER_DELAY -WFG_START_DELAY); sim3shaped_pulse("","offC9","",0.0,pwC8, 2.0*pwN, zero, zero, zero, 0.0, 0.0); } else sim3shaped_pulse("","offC9","",0.0,pwC8, 2.0*pwN, zero, zero, zero, 0.0, 0.0); delay(timeTC- 2.0e-6 -WFG_STOP_DELAY-POWER_DELAY); decpwrf(rf2); decrgpulse(pwC2, zero, 2.0e-6, 2.0e-6); /* 13Ca 180 degree pulse */ delay(timeTC-tau1- 4.0e-6 -SAPS_DELAY); phshift9 = 230.0; /* = 320-90 - correction for -90 degree phase shift in F1 */ initval(phshift9, v9); decstepsize(1.0); dcplrphase(v9); /* SAPS_DELAY */ } else /* xxxxxxx 13Ca Conventional EVOLUTION xxxxxxxxx */ { if ((ni>1.0) && (tau1>0.0)) /* total 13C evolution equals d2 exactly */ { /* 2.0*pwC1/PI compensates for evolution at 64% rate during pwC1 */ decpwrf(rf9); if(tau1 - 2.0*pwC1/PI - WFG3_START_DELAY - 0.5*pwZ > 0.0) { delay(tau1 - 2.0*pwC1/PI - WFG3_START_DELAY - 0.5*pwZ); sim3shaped_pulse("", "offC9", "", 0.0, pwC8, 2.0*pwN, zero, zero, zero, 0.0, 0.0); initval(phshift9, v9); decstepsize(1.0); dcplrphase(v9); /* SAPS_DELAY */ delay(tau1 - 2.0*pwC1/PI - SAPS_DELAY - 0.5*pwZ - 2.0e-6); } else { initval(180.0, v9); decstepsize(1.0); dcplrphase(v9); /* SAPS_DELAY */ delay(2.0*tau1 - 4.0*pwC1/PI - SAPS_DELAY - 2.0e-6); } } else /* 13Ca evolution refocused for 1st increment */ { decpwrf(rf2); delay(10.0e-6); decrgpulse(pwC2, zero, 2.0e-6, 0.0); delay(10.0e-6); } } decphase(t5); decpwrf(rf1); decrgpulse(pwC1, t5, 2.0e-6, 0.0); /* xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx */ xmtroff(); obsprgoff(); rgpulse(pwHd,three,2.0e-6,0.0); decoffset(dofCO); decpwrf(rf6); decphase(one); if ( dm3[B] == 'y' ) /* turns off 2H decoupling */ { setstatus(DEC3ch, FALSE, 'c', FALSE, dmf3); dec3rgpulse(1/dmf3,three,2.0e-6,2.0e-6); dec3blank(); lk_autotrig(); /* resumes lock pulsing */ } zgradpulse(gzlvl10, gt10); delay(2.0e-4); decshaped_pulse("offC6", pwC6, one, 0.0, 0.0); zgradpulse(gzlvl8, gt7); decpwrf(rf0); decphase(zero); delay(tauC - gt7 - 0.5*10.933*pwC); decrgpulse(pwC*158.0/90.0, zero, 0.0, 0.0); decrgpulse(pwC*171.2/90.0, two, 0.0, 0.0); decrgpulse(pwC*342.8/90.0, zero, 0.0, 0.0); /* Shaka 6 composite */ decrgpulse(pwC*145.5/90.0, two, 0.0, 0.0); decrgpulse(pwC*81.2/90.0, zero, 0.0, 0.0); decrgpulse(pwC*85.3/90.0, two, 0.0, 0.0); zgradpulse(gzlvl8, gt7); decpwrf(rf6); decphase(zero); delay(tauC - gt7 - 0.5*10.933*pwC - WFG_START_DELAY); /* WFG_START_DELAY */ decshaped_pulse("offC6", pwC6, zero, 0.0, 0.0); /* xxxxxxxxxxxxxxxxxx OPTIONS FOR N15 EVOLUTION xxxxxxxxxxxxxxxxxxxxx */ zgradpulse(gzlvl4, gt4); txphase(one); decphase(zero); decpwrf(rf8); dcplrphase(zero); dec2phase(t8); delay(2.0e-4); if (TROSY[A]=='n') {rgpulse(pwHd,one,0.0,0.0); txphase(zero); delay(2.0e-6); obsprgon("waltz16", pwHd, 90.0); xmtron();} dec2rgpulse(pwN, t8, 0.0, 0.0); /* N15 EVOLUTION BEGINS HERE */ dec2phase(t9); if(SCT[A] == 'y') { delay(t2a); dec2rgpulse(2.0*pwN, t9, 0.0, 0.0); delay(t2b); decshaped_pulse("offC8", pwC8, zero, 0.0, 0.0); /* WFG_START_DELAY */ } else { delay(timeTN - WFG3_START_DELAY - tau2); /* WFG3_START_DELAY */ sim3shaped_pulse("", "offC8", "", 0.0, pwC8, 2.0*pwN, zero, zero, t9, 0.0, 0.0); } dec2phase(t10); decpwrf(rf3); if (TROSY[A]=='y') { if (tau2 > gt1 + 2.0*GRADIENT_DELAY + 1.5e-4 + pwHs) { txphase(three); delay(timeTN - pwC3 - WFG_START_DELAY); /* WFG_START_DELAY */ decshaped_pulse("offC3", pwC3, zero, 0.0, 0.0); delay(tau2 - gt1 - 2.0*GRADIENT_DELAY - 1.5e-4 - pwHs); if (mag_flg[A]=='y') magradpulse(gzcal*gzlvl1, gt1); else zgradpulse(gzlvl1, gt1); /* 2.0*GRADIENT_DELAY */ obspower(tpwrs); /* POWER_DELAY */ delay(1.0e-4 - POWER_DELAY); if (tpwrsf < 4095.0) {obspwrf(tpwrsf); obspower(tpwrs+6.0);} /* increases tpwrs by 6dB, now need */ else obspower(tpwrs); /* tpwrsf to be ~ 2048 for equivalence */ shaped_pulse("H2Osinc", pwHs, three, 0.0, 0.0); obspower(tpwr); obspwrf(4095.0); txphase(t4); delay(0.5e-4 - POWER_DELAY); } else if (tau2 > pwHs + 0.5e-4) { txphase(three); delay(timeTN-pwC3-WFG_START_DELAY-gt1-2.0*GRADIENT_DELAY-1.0e-4); if (mag_flg[A]=='y') magradpulse(gzcal*gzlvl1, gt1); else zgradpulse(gzlvl1, gt1); /* 2.0*GRADIENT_DELAY */ obspower(tpwrs); /* POWER_DELAY */ delay(1.0e-4 - POWER_DELAY); /* WFG_START_DELAY */ decshaped_pulse("offC3", pwC3, zero, 0.0, 0.0); delay(tau2 - pwHs - 0.5e-4); if (tpwrsf < 4095.0) {obspwrf(tpwrsf); obspower(tpwrs+6.0);} /* increases tpwrs by 6dB, now need */ else obspower(tpwrs); /* tpwrsf to be ~ 2048 for equivalence */ shaped_pulse("H2Osinc", pwHs, three, 0.0, 0.0); obspower(tpwr); obspwrf(4095.0); txphase(t4); delay(0.5e-4 - POWER_DELAY); } else { txphase(three); delay(timeTN - pwC3 - WFG_START_DELAY - gt1 - 2.0*GRADIENT_DELAY - 1.5e-4 - pwHs); if (mag_flg[A]=='y') magradpulse(gzcal*gzlvl1, gt1); else zgradpulse(gzlvl1, gt1); /* 2.0*GRADIENT_DELAY */ obspower(tpwrs); /* POWER_DELAY */ delay(1.0e-4 - POWER_DELAY); /* WFG_START_DELAY */ if (tpwrsf < 4095.0) {obspwrf(tpwrsf); obspower(tpwrs+6.0);} /* increases tpwrs by 6dB, now need */ else obspower(tpwrs); /* tpwrsf to be ~ 2048 for equivalence */ shaped_pulse("H2Osinc", pwHs, three, 0.0, 0.0); obspower(tpwr); obspwrf(4095.0); txphase(t4); delay(0.5e-4 - POWER_DELAY); decshaped_pulse("offC3", pwC3, zero, 0.0, 0.0); delay(tau2); } } else { if (tau2 > kappa) { delay(timeTN - pwC3 - WFG_START_DELAY); /* WFG_START_DELAY */ decshaped_pulse("offC3", pwC3, zero, 0.0, 0.0); delay(tau2 - kappa - PRG_STOP_DELAY - pwHd - 2.0e-6); xmtroff(); obsprgoff(); /* PRG_STOP_DELAY */ rgpulse(pwHd,three,2.0e-6,0.0); txphase(t4); delay(kappa - gt1 - 2.0*GRADIENT_DELAY - 1.0e-4); if (mag_flg[A]=='y') magradpulse(gzcal*gzlvl1, gt1); else zgradpulse(gzlvl1, gt1); /* 2.0*GRADIENT_DELAY */ obspower(tpwr); /* POWER_DELAY */ delay(1.0e-4 - POWER_DELAY); } else if (tau2 > (kappa - pwC3 - WFG_START_DELAY)) { delay(timeTN + tau2 - kappa - PRG_STOP_DELAY - pwHd - 2.0e-6); xmtroff(); obsprgoff(); /* PRG_STOP_DELAY */ rgpulse(pwHd,three,2.0e-6,0.0); txphase(t4); /* WFG_START_DELAY */ decshaped_pulse("offC3", pwC3, zero, 0.0, 0.0); delay(kappa -pwC3 -WFG_START_DELAY -gt1 -2.0*GRADIENT_DELAY -1.0e-4); if (mag_flg[A]=='y') magradpulse(gzcal*gzlvl1, gt1); else zgradpulse(gzlvl1, gt1); /* 2.0*GRADIENT_DELAY */ obspower(tpwr); /* POWER_DELAY */ delay(1.0e-4 - POWER_DELAY); } else if (tau2 > gt1 + 2.0*GRADIENT_DELAY + 1.0e-4) { delay(timeTN + tau2 - kappa - PRG_STOP_DELAY - pwHd - 2.0e-6); xmtroff(); obsprgoff(); /* PRG_STOP_DELAY */ rgpulse(pwHd,three,2.0e-6,0.0); txphase(t4); delay(kappa - tau2 - pwC3 - WFG_START_DELAY); /* WFG_START_DELAY */ decshaped_pulse("offC3", pwC3, zero, 0.0, 0.0); delay(tau2 - gt1 - 2.0*GRADIENT_DELAY - 1.0e-4); if (mag_flg[A]=='y') magradpulse(gzcal*gzlvl1, gt1); else zgradpulse(gzlvl1, gt1); /* 2.0*GRADIENT_DELAY */ obspower(tpwr); /* POWER_DELAY */ delay(1.0e-4 - POWER_DELAY); } else { delay(timeTN + tau2 - kappa - PRG_STOP_DELAY - pwHd - 2.0e-6); xmtroff(); obsprgoff(); /* PRG_STOP_DELAY */ rgpulse(pwHd,three,2.0e-6,0.0); txphase(t4); delay(kappa-tau2-pwC3-WFG_START_DELAY-gt1-2.0*GRADIENT_DELAY-1.0e-4); if (mag_flg[A]=='y') magradpulse(gzcal*gzlvl1, gt1); else zgradpulse(gzlvl1, gt1); /* 2.0*GRADIENT_DELAY */ obspower(tpwr); /* POWER_DELAY */ delay(1.0e-4 - POWER_DELAY); /* WFG_START_DELAY */ decshaped_pulse("offC3", pwC3, zero, 0.0, 0.0); delay(tau2); } } /* xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx */ if (TROSY[A]=='y') rgpulse(pw, t4, 0.0, 0.0); else sim3pulse(pw, 0.0, pwN, t4, zero, t10, 0.0, 0.0); txphase(zero); dec2phase(zero); zgradpulse(gzlvl5, gt5); if (TROSY[A]=='y') delay(lambda - 0.65*(pw + pwN) - gt5); else delay(lambda - 1.3*pwN - gt5); sim3pulse(2.0*pw, 0.0, 2.0*pwN, zero, zero, zero, 0.0, 0.0); zgradpulse(gzlvl5, gt5); txphase(one); dec2phase(t11); delay(lambda - 1.3*pwN - gt5); sim3pulse(pw, 0.0, pwN, one, zero, t11, 0.0, 0.0); txphase(zero); dec2phase(zero); zgradpulse(gzlvl6, gt5); delay(lambda - 1.3*pwN - gt5); sim3pulse(2.0*pw, 0.0, 2.0*pwN, zero, zero, zero, 0.0, 0.0); dec2phase(t10); zgradpulse(gzlvl6, gt5); if (TROSY[A]=='y') delay(lambda - 1.6*pwN - gt5); else delay(lambda - 0.65*pwN - gt5); if (TROSY[A]=='y') dec2rgpulse(pwN, t10, 0.0, 0.0); else rgpulse(pw, zero, 0.0, 0.0); delay((gt1/10.0) + 1.0e-4 +gstab - 0.5*pw + 2.0*GRADIENT_DELAY + POWER_DELAY); rgpulse(2.0*pw, zero, 0.0,rof1); dec2power(dpwr2); /* POWER_DELAY */ if (mag_flg[A] == 'y') magradpulse(icosel*gzcal*gzlvl2, gt1/10.0); else zgradpulse(icosel*gzlvl2, gt1/10.0); /* 2.0*GRADIENT_DELAY */ delay(gstab); rcvron(); statusdelay(C,1.0e-4 - rof1); if (dm3[B]=='y') lk_sample(); setreceiver(t12); }

void pulsesequence() { /* DECLARE VARIABLES */ char shape_ss[MAXSTR]; int t1_counter; double tau1, /* t1/2 */ taua = getval("taua"), /* 2.25ms */ taub = getval("taub"), /* 2.75ms */ time_T1, pwN, /* PW90 for N-nuc */ pwNlvl, /* power level for N hard pulses */ ncyc = getval("ncyc"), compH= getval("compH"), pwHs = getval("pwHs"), /* H1 90 degree pulse length at tpwrs */ tpwrs , /* power for the pwHs ("H2Osinc") pulse */ tpwrsf , /* fine power for the pwHs ("H2Osinc") pulse */ shss_pwr, /* power for cos modulated NH pulses */ pw_shpss=getval("pw_shpss"), waterdly, /* pw for water pulse */ waterpwrf, /* fine power for water pulse */ waterpwr, /* power for water pulse */ gt0, gt1 = getval("gt1"), gt2 = getval("gt2"), gt3 = getval("gt3"), gt4 = getval("gt4"), gt5 = getval("gt5"), gt6 = getval("gt6"), gzlvl0 = getval("gzlvl0"), gzlvl1 = getval("gzlvl1"), gzlvl2 = getval("gzlvl2"), gzlvl3 = getval("gzlvl3"), gzlvl4 = getval("gzlvl4"), gzlvl5 = getval("gzlvl5"), gzlvl6 = getval("gzlvl6"); /* LOAD VARIABLES */ pwN = getval("pwN"); pwNlvl = getval("pwNlvl"); tpwrsf = getval("tpwrsf"); waterpwrf = getval("waterpwrf"); waterdly = getval("waterdly"); getstr("shape_ss",shape_ss); time_T1=ncyc*(2.0*2.5e-3+pw_shpss); if (ix==1) printf(" ncyc= %f, time_T1= %f \n", ncyc,time_T1); /* selective H20 one-lobe sinc pulse */ tpwrs = tpwr - 20.0*log10(pwHs/(compH*pw*1.69)); /* needs 1.69 times more */ tpwrs = (int) (tpwrs); /* power than a square pulse */ /* selective H20 watergate pulse */ waterpwr = tpwr - 20.0*log10(waterdly/(compH*pw)); waterpwr = (int) (waterpwr); /* selective cos modulated NH 180 degree pulse */ shss_pwr = tpwr - 20.0*log10(pw_shpss/((compH*2*pw)*2)); /* needs 2 times more */ shss_pwr = (int) (shss_pwr); /* power than a square pulse */ /* check validity of parameter range */ if((dm[A] == 'y' || dm[B] == 'y' )) { printf("incorrect Dec1 decoupler flags! "); psg_abort(1); } if (dm2[A] == 'y' || dm2[B] == 'y' || dm2[C] == 'y') { printf("incorrect Dec2 decoupler flag! dm2 should be 'nnn' "); psg_abort(1); } if (dmm2[A] == 'g' || dmm2[B] == 'g' || dmm2[C] == 'g') { printf("incorrect Dec2 decoupler flag! dmm2 should be 'ccc' "); psg_abort(1); } if( dpwr > 50 ) { printf("don't fry the probe, dpwr too large! "); psg_abort(1); } if( dpwr2 > 50 ) { printf("don't fry the probe, dpwr2 too large! "); psg_abort(1); } if(gt1 > 15.0e-3 || gt2 > 15.0e-3 || gt3 > 15.0e-3 || gt4 > 15.0e-3) { printf("gti must be less than 15 ms \n"); psg_abort(1); } /* LOAD VARIABLES */ settable(t1, 8, phi1); settable(t2, 4, phi2); settable(t3, 1, phi3); settable(t10, 8, phi10); settable(t14, 8, rec); /* Phase incrementation for hypercomplex data */ if ( phase1 == 2 ) /* Hypercomplex in t1 */ { ttadd(t14,t10,4); tsadd(t3,2,4); } /* calculate modification to phases based on current t1 values to achieve States-TPPI acquisition */ if(ix == 1) d2_init = d2; t1_counter = (int) ( (d2-d2_init)*sw1 + 0.5); tau1=0.5*d2; if(t1_counter %2) { tsadd(t2,2,4); tsadd(t14,2,4); } /* BEGIN ACTUAL PULSE SEQUENCE */ status(A); obspower(tpwr); /* Set power for pulses */ dec2power(pwNlvl); /* Set decoupler2 power to pwNlvl */ initval(ncyc+0.1,v10); /* for DIPSI-2 */ delay(d1); status(B); rcvroff(); /*destroy N15 magnetization*/ dec2rgpulse(pwN, zero, 0.0, 0.0); zgradpulse(gzlvl1, gt1); delay(9.0e-5); /* 1H-15N INEPT */ rgpulse(pw,zero,1.0e-6,0.0); txphase(zero); dec2phase(zero); zgradpulse(gzlvl2,gt2); delay(taua -pwN-0.5*pw -gt2 ); /* delay=1/4J(NH) */ sim3pulse(2.0*pw,0.0e-6,2*pwN,zero,zero,zero,0.0,0.0); txphase(one); dec2phase(t1); zgradpulse(gzlvl2,gt2); delay(taua -1.5*pwN -gt2); /* delay=1/4J(NH) */ sim3pulse(pw,0.0e-6,pwN,one,zero,t1,0.0,0.0); if (tpwrsf < 4095.0) {obspwrf(tpwrsf); tpwrs=tpwrs+6.0;} obspower(tpwrs); shaped_pulse("H2Osinc", pwHs, two, 2.0e-6, 0.0); obspower(tpwr); obspwrf(4095.0); tpwrs=tpwrs-6.0; txphase(zero); dec2phase(zero); zgradpulse(gzlvl3,gt3); delay(taub -1.5*pwN -gt3 -pwHs-2.0e-6-2.0*POWER_DELAY-WFG_START_DELAY); /* delay=1/4J(NH) */ sim3pulse(2.0*pw,0.0e-6,2*pwN,zero,zero,zero,0.0,0.0); dec2phase(one); zgradpulse(gzlvl3,gt3); delay(taub -1.5*pwN -gt3 ); /* delay=1/4J(NH) */ dec2phase(one); dec2rgpulse(pwN,one,0.0,0.0); /* relaxation recovery */ if (ncyc>0.6) { obspower(shss_pwr); starthardloop(v10); delay(2.5e-3); shaped_pulse(shape_ss,pw_shpss,zero,0.0,0.0); delay(2.5e-3); endhardloop(); obspower(tpwr); } zgradpulse(gzlvl6,gt6); delay(200.0e-6); dec2rgpulse(pwN,t2,0.0,0.0); txphase(t3); dec2phase(zero); /* evolution of t1 */ if(d2>0.001) { zgradpulse( gzlvl0,(d2/2.0-0.0003-2.0*GRADIENT_DELAY)); delay(300.0e-6); zgradpulse(-gzlvl0,(d2/2.0-0.0003-2.0*GRADIENT_DELAY)); delay(300.0e-6); } else delay(d2); /* ST2 */ rgpulse(pw,t3,0.0,0.0); txphase(t3); if (waterpwrf < 4095.0) {obspwrf(waterpwrf); waterpwr=waterpwr+6.0;} obspower(waterpwr); rgpulse(waterdly,t3,0.0,rof1); if (waterpwrf < 4095.0) {obspwrf(4095.0); waterpwr=waterpwr-6.0;} obspower(tpwr); txphase(zero); zgradpulse(gzlvl4,gt4); delay(taua -pwN -0.5*pw -gt4-waterdly-rof1); sim3pulse(2.0*pw,0.0e-6,2*pwN,zero,zero,zero,0.0,0.0); dec2phase(t3); zgradpulse(gzlvl4,gt4); delay(taua -1.5*pwN -gt4 -waterdly-rof1); /* delay=1/4J(NH) */ if (waterpwrf < 4095.0) {obspwrf(waterpwrf); waterpwr=waterpwr+6.0;} obspower(waterpwr); txphase(two); rgpulse(waterdly,two,rof1,0.0); if (waterpwrf < 4095.0) {obspwrf(4095.0); waterpwr=waterpwr-6.0;} obspower(tpwr); sim3pulse(pw,0.0e-6,pwN,zero,zero,t3,0.0,0.0); /* watergate */ zgradpulse(gzlvl5,gt5); delay(taua-1.5*pwN-waterdly-gt5); txphase(two); if (waterpwrf < 4095.0) {obspwrf(waterpwrf); waterpwr=waterpwr+6.0;} obspower(waterpwr); dec2phase(zero); rgpulse(waterdly,two,0.0,rof1); if (waterpwrf < 4095.0) {obspwrf(4095.0); waterpwr=waterpwr-6.0;} obspower(tpwr); txphase(zero); sim3pulse(2.0*pw,0.0e-6,2*pwN,zero,zero,zero,0.0,0.0); if (waterpwrf < 4095.0) {obspwrf(waterpwrf); waterpwr=waterpwr+6.0;} obspower(waterpwr); txphase(two); rgpulse(waterdly,two,rof1,0.0); if (waterpwrf < 4095.0) {obspwrf(4095.0); waterpwr=waterpwr-6.0;} zgradpulse(gzlvl5,gt5); obspwrf(4095.0); obspower(tpwr); delay(taua-1.5*pwN-waterdly-gt5); dec2rgpulse(pwN,zero,0.0,0.0); /* acquire data */ status(C); setreceiver(t14); }

pulsesequence() { /* Internal variable declarations *********************/ char txphase[MAXSTR]; char rxphase[MAXSTR]; char blankmode[MAXSTR]={0}; double postDelay; double rfDuration; double acqt; int i,ret=-1; static int phs1[4] = {0,2,1,3}; /* from T1meas.c */ /*************************************************/ /* Initialize paramter **************************/ i = 0; postDelay = 0.5; acqt = 0.0; getstr("rxphase",rxphase); getstr("txphase",txphase); ret = P_getstring(GLOBAL,"blankmode",blankmode,1,MAXSTR); //getparm("blankmode","string",GLOBAL,blankmode,MAXSTR); postDelay = tr - at; //printf("blankmode=%s\n",blankmode); /*************************************************/ /* check phase setting ***************************/ if ( (txphase[0] =='n') && (rxphase[0] =='n') ) { abort_message("ERROR - Select at least one phase [Tx or Rx]\n"); } /**************************************************/ /* check pulse width *****************************/ rfDuration = shapelistpw(p1pat, p1); /* assign exitation pulse duration */ acqt = rfDuration + rof1 - alfa; if (FP_GT(acqt, at)) { abort_message("Pulse duration too long. max [%.3f] ms\n",(at-rof1+alfa)*1000.0); } if(ret==0 && blankmode[0]=='u') obsunblank(); delay(postDelay); settable(t1,4,phs1); /*from T1meas.c */ getelem(t1,ct,v11); /*from T1meas.c */ setreceiver(t1); /*==============================================*/ /* START LOOPBACK PULSE SEQUENCE */ /*==============================================*/ status(A); obsoffset(resto); /* TTL trigger to scope sequence ****************************/ sp1on(); /* Relaxation delay ***********************************/ xgate(ticks); /* RF pulse *******************************************/ obspower(tpwr); obspwrf(tpwrf); ShapedXmtNAcquire(p1pat, rfDuration, v11, rof1, OBSch); endacq(); sp1off(); if(ret==0 && blankmode[0]=='u') obsunblank(); }

pulsesequence() { /* DECLARE AND LOAD VARIABLES */ char f1180[MAXSTR], /* Flag to start t1 @ halfdwell */ mag_flg[MAXSTR], /*magic-angle coherence transfer gradients */ f2180[MAXSTR], /* Flag to start t2 @ halfdwell */ C13refoc[MAXSTR], /* C13 sech/tanh pulse in middle of t1*/ NH2only[MAXSTR]; /* spectrum of only NH2 groups */ int icosel, /* used to get n and p type */ t1_counter, /* used for states tppi in t1 */ t2_counter, /* used for states tppi in t2 */ PRexp, /* projection-reconstruction flag */ ni2 = getval("ni2"); double tau1, /* t1 delay */ mix = getval("mix"), /* NOESY mix time */ tau2, /* t2 delay */ lambda = 0.94/(4.0*getval("JNH")), /* 1/4J H1 evolution delay */ tNH = 1.0/(4.0*getval("JNH")), /* 1/4J N15 evolution delay */ csa, sna, pra = M_PI*getval("pra")/180.0, /* temporary Pbox parameters */ bw, pws, ofs, ppm, nst, /* bandwidth, pulsewidth, offset, ppm, # steps */ /* the sech/tanh pulse is automatically calculated by the macro "biocal", */ /* and is called directly from your shapelib. */ pwClvl = getval("pwClvl"), /* coarse power for C13 pulse */ pwC = getval("pwC"), /* C13 90 degree pulse length at pwClvl */ rf0, /* maximum fine power when using pwC pulses */ rfst, /* fine power for the stCall pulse */ compH = getval("compH"), /* adjustment for H1 amplifier compression */ compC = getval("compC"), /* adjustment for C13 amplifier compression */ dof100, /* C13 frequency at 100ppm for both aliphatic & aromatic*/ tpwrsf = getval("tpwrsf"), /* fine power adjustment for flipback pulse*/ pwHs = getval("pwHs"), /* H1 90 degree pulse length at tpwrs */ tpwrs, /* power for the pwHs ("H2Osinc") pulse */ pwNlvl = getval("pwNlvl"), /* power for N15 pulses */ pwN = getval("pwN"), /* N15 90 degree pulse length at pwNlvl */ sw1 = getval("sw1"), sw2 = getval("sw2"), gzcal=getval("gzcal"), gt1 = getval("gt1"), /* coherence pathway gradients */ gzlvl1 = getval("gzlvl1"), gzlvl2 = getval("gzlvl2"), gt0 = getval("gt0"), /* other gradients */ gt3 = getval("gt3"), gt4 = getval("gt4"), gt5 = getval("gt5"), gstab = getval("gstab"), gzlvl0 = getval("gzlvl0"), gzlvl3 = getval("gzlvl3"), gzlvl6 = getval("gzlvl6"), gzlvl4 = getval("gzlvl4"), gzlvl5 = getval("gzlvl5"); getstr("f1180",f1180); getstr("mag_flg",mag_flg); getstr("f2180",f2180); getstr("C13refoc",C13refoc); getstr("NH2only",NH2only); csa = cos(pra); sna = sin(pra); /* LOAD PHASE TABLE */ settable(t1,2,phi1); settable(t3,4,phi3); settable(t9,16,phi9); settable(t10,1,phi10); settable(t11,8,rec); /* INITIALIZE VARIABLES */ /* maximum fine power for pwC pulses (and initialize rfst) */ rf0 = 4095.0; rfst=0.0; setautocal(); /* activate auto-calibration flags */ if (autocal[0] == 'n') { /* 180 degree adiabatic C13 pulse from 0 to 200 ppm */ if (C13refoc[A]=='y') { rfst = (compC*4095.0*pwC*4000.0*sqrt((30.0*sfrq/600.0+7.0)/0.35)); rfst = (int) (rfst + 0.5); if ( 1.0/(4000.0*sqrt((30.0*sfrq/600.0+7.0)/0.35)) < pwC ) { text_error( " Not enough C13 RF. pwC must be %f usec or less.\n", (1.0e6/(4000.0*sqrt((30.0*sfrq/600.0+7.0)/0.35))) ); psg_abort(1); } } } else /* if autocal = 'y'(yes), 'q'(quiet), r(read), or 's'(semi) */ { if(FIRST_FID) /* call Pbox */ { if (C13refoc[A]=='y') { ppm = getval("dfrq"); ofs = 0.0; pws = 0.001; /* 1 ms long pulse */ bw = 200.0*ppm; nst = 1000; /* nst - number of steps */ stC200 = pbox_makeA("stC200", "sech", bw, pws, ofs, compC*pwC, pwClvl, nst); } ofs_check(H1ofs, C13ofs, N15ofs, H2ofs); } if (C13refoc[A]=='y') rfst = stC200.pwrf; } /* 180 degree adiabatic C13 pulse from 0 to 200 ppm */ dof100 = dof + 65.0*dfrq; /* selective H20 one-lobe sinc pulse */ tpwrs = tpwr - 20.0*log10(pwHs/((compH*pw)*1.69)); /* needs 1.69 times more */ tpwrs = (int) (tpwrs); /* power than a square pulse */ /* CHECK VALIDITY OF PARAMETER RANGES */ if ( (mix - gt4 - gt5) < 0.0 ) { text_error("mix is too small. Make mix equal to %f or more.\n",(gt4 + gt5)); psg_abort(1); } if((dm[A] == 'y' || dm[B] == 'y' || dm[C] == 'y' )) { text_error("incorrect dec1 decoupler flags! Should be 'nnn' "); psg_abort(1); } if((dm2[A] == 'y' || dm2[B] == 'y')) { text_error("incorrect dec2 decoupler flags! Should be 'nny' "); psg_abort(1); } if( dpwr2 > 50 ) { text_error("don't fry the probe, DPWR2 too large! "); psg_abort(1); } if( pw > 20.0e-6 ) { text_error("dont fry the probe, pw too high ! "); psg_abort(1); } if( pwN > 100.0e-6 ) { text_error("dont fry the probe, pwN too high ! "); psg_abort(1); } /* PHASES AND INCREMENTED TIMES */ /* Phase incrementation for hypercomplex 2D data, States-Haberkorn element */ if (phase1 == 2) tsadd(t1,1,4); if (phase2 == 1) { tsadd(t10,2,4); icosel = 1; } else icosel = -1; /* Set up f1180 */ PRexp = 0; if((pra > 0.0) && (pra < 90.0)) PRexp = 1; if(PRexp) /* set up Projection-Reconstruction experiment */ tau1 = d2*csa; else tau1 = d2; if((f1180[A] == 'y') && (ni > 1.0)) { tau1 += ( 1.0 / (2.0*sw1) ); if(tau1 < 0.2e-6) tau1 = 0.0; } tau1 = tau1/2.0; /* Set up f2180 */ if(PRexp) tau2 = d2*sna; else { tau2 = d3; if((f2180[A] == 'y') && (ni2 > 1.0)) { tau2 += ( 1.0 / (2.0*sw2) ); if(tau2 < 0.2e-6) tau2 = 0.0; } } tau2 = tau2/2.0; /* Calculate modifications to phases for States-TPPI acquisition */ if( ix == 1) d2_init = d2; t1_counter = (int) ( (d2-d2_init)*sw1 + 0.5 ); if(t1_counter % 2) { tsadd(t1,2,4); tsadd(t11,2,4); } if( ix == 1) d3_init = d3; t2_counter = (int) ( (d3-d3_init)*sw2 + 0.5 ); if(t2_counter % 2) { tsadd(t3,2,4); tsadd(t11,2,4); } /* Correct inverted signals for NH2 only spectra */ if (NH2only[A]=='y') { tsadd(t3,2,4); } /* BEGIN PULSE SEQUENCE */ status(A); obspower(tpwr); decpower(pwClvl); dec2power(pwNlvl); decoffset(dof); decpwrf(rf0); txphase(zero); dec2phase(zero); delay(d1); dec2rgpulse(pwN, zero, 0.0, 0.0); /*destroy N15 and C13 magnetization*/ decrgpulse(pwC, zero, 0.0, 0.0); zgradpulse(gzlvl0, 0.5e-3); delay(1.0e-4); dec2rgpulse(pwN, one, 0.0, 0.0); decrgpulse(pwC, one, 0.0, 0.0); zgradpulse(0.7*gzlvl0, 0.5e-3); txphase(t1); decphase(zero); dec2phase(zero); initval(135.0,v1); obsstepsize(1.0); xmtrphase(v1); delay(5.0e-4); rcvroff(); rgpulse(pw, t1, 50.0e-6, 0.0); /* 1H pulse excitation */ xmtrphase(zero); /* SAPS_DELAY */ txphase(zero); if (tau1 > (2.0*GRADIENT_DELAY + pwN + 0.64*pw + 5.0*SAPS_DELAY)) { if (tau1>0.002) { zgradpulse(gzlvl6, 0.8*(tau1 - 2.0*GRADIENT_DELAY - pwN - 0.64*pw)); delay(0.2*(tau1 - 2.0*GRADIENT_DELAY - pwN - 0.64*pw) - SAPS_DELAY); } else { delay(tau1-pwN-0.64*pw); } if (C13refoc[A]=='y') sim3pulse(0.0, 2.0*pwC, 2.0*pwN, zero, zero, zero, 0.0, 0.0); else dec2rgpulse(2.0*pwN, zero, 0.0, 0.0); if (tau1>0.002) { zgradpulse(-1.0*gzlvl6, 0.8*(tau1 - 2.0*GRADIENT_DELAY - pwN - 0.64*pw)); delay(0.2*(tau1 - 2.0*GRADIENT_DELAY - pwN - 0.64*pw) - SAPS_DELAY); } else { delay(tau1-pwN-0.64*pw); } } else if (tau1 > (0.64*pw + 0.5*SAPS_DELAY)) delay(2.0*tau1 - 2.0*0.64*pw - SAPS_DELAY ); rgpulse(pw, zero, 0.0, 0.0); delay(mix - gt4 - gt5 -gstab -200.0e-6); dec2rgpulse(pwN, zero, 0.0, 0.0); zgradpulse(gzlvl4, gt4); delay(gstab); rgpulse(pw, zero, 200.0e-6,0.0); /* HSQC begins */ dec2phase(zero); zgradpulse(gzlvl0, gt0); delay(lambda - gt0); sim3pulse(2.0*pw, 0.0, 2.0*pwN, zero, zero, zero, 0.0, 0.0); txphase(one); zgradpulse(gzlvl0, gt0); delay(lambda - gt0); rgpulse(pw, one, 0.0, 0.0); txphase(two); if (tpwrsf<4095.0) { obspower(tpwrs+6.0); obspwrf(tpwrsf); shaped_pulse("H2Osinc", pwHs, two, 5.0e-4, 0.0); obspower(tpwr); obspwrf(4095.0); } else { obspower(tpwrs); shaped_pulse("H2Osinc", pwHs, two, 5.0e-4, 0.0); obspower(tpwr); } zgradpulse(gzlvl3, gt3); dec2phase(t3); decpwrf(rfst); decoffset(dof100); delay(2.0e-4); dec2rgpulse(pwN, t3, 0.0, 0.0); decphase(zero); /* xxxxxxxxxxxxxxxxxx OPTIONS FOR N15 EVOLUTION xxxxxxxxxxxxxxxxxxxxx */ txphase(zero); dec2phase(t9); if (NH2only[A]=='y') { delay(tau2); /* optional sech/tanh pulse in middle of t2 */ if (C13refoc[A]=='y') /* WFG_START_DELAY */ { decshaped_pulse("stC200", 1.0e-3, zero, 0.0, 0.0); delay(tNH - 1.0e-3 - WFG_START_DELAY - 2.0*pw); } else { delay(tNH - 2.0*pw); } rgpulse(2.0*pw, zero, 0.0, 0.0); if (tNH < gt1 + 1.99e-4) delay(gt1 + 1.99e-4 - tNH); delay(tau2); dec2rgpulse(2.0*pwN, t9, 0.0, 0.0); if (mag_flg[A] == 'y') { magradpulse(gzcal*gzlvl1, gt1); } else { zgradpulse(gzlvl1, gt1); } dec2phase(t10); if (tNH > gt1 + 1.99e-4) delay(tNH - gt1 - 2.0*GRADIENT_DELAY); else delay(1.99e-4 - 2.0*GRADIENT_DELAY); } else { if ( (C13refoc[A]=='y') && (tau2 > 0.5e-3 + WFG2_START_DELAY) ) { delay(tau2 - 0.5e-3 - WFG2_START_DELAY); /* WFG2_START_DELAY */ simshaped_pulse("", "stC200", 2.0*pw, 1.0e-3, zero, zero, 0.0, 0.0); delay(tau2 - 0.5e-3); delay(gt1 + 2.0e-4); } else { delay(tau2); rgpulse(2.0*pw, zero, 0.0, 0.0); delay(gt1 + 2.0e-4 - 2.0*pw); delay(tau2); } dec2rgpulse(2.0*pwN, t9, 0.0, 0.0); if (mag_flg[A] == 'y') { magradpulse(gzcal*gzlvl1, gt1); } else { zgradpulse(gzlvl1, gt1); } dec2phase(t10); delay(2.0e-4 - 2.0*GRADIENT_DELAY); } /* xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx */ sim3pulse(pw, 0.0, pwN, zero, zero, t10, 0.0, 0.0); dec2phase(zero); zgradpulse(gzlvl5, gt5); delay(lambda - 1.5*pwN - gt5); sim3pulse(2.0*pw, 0.0, 2.0*pwN, zero, zero, zero, 0.0, 0.0); zgradpulse(gzlvl5, gt5); txphase(one); dec2phase(one); delay(lambda - 1.5*pwN - gt5); sim3pulse(pw, 0.0, pwN, one, zero, one, 0.0, 0.0); txphase(zero); dec2phase(zero); zgradpulse(1.5*gzlvl5, gt5); delay(lambda - 1.5*pwN - gt5); sim3pulse(2.0*pw, 0.0, 2.0*pwN, zero, zero, zero, 0.0, 0.0); zgradpulse(1.5*gzlvl5, gt5); delay(lambda - pwN - 0.5*pw - gt5); rgpulse(pw, zero, 0.0, 0.0); delay((gt1/10.0) + 1.0e-4+ gstab - 0.5*pw + 2.0*GRADIENT_DELAY + POWER_DELAY); rgpulse(2.0*pw, zero, 0.0, rof1); dec2power(dpwr2); /* POWER_DELAY */ if (mag_flg[A] == 'y') { magradpulse(icosel*gzcal*gzlvl2, gt1/10.0); } else { zgradpulse(icosel*gzlvl2, gt1/10.0); } delay(gstab); rcvron(); statusdelay(C,1.0e-4-rof1); setreceiver(t11); }

pulsesequence() { /* DECLARE VARIABLES */ char satmode[MAXSTR], fscuba[MAXSTR], fc180[MAXSTR], /* Flag for checking sequence */ ddseq[MAXSTR], /* 2H decoupling seqfile */ fCTCa[MAXSTR], /* Flag for CT or non_CT on Ca dimension */ sel_flg[MAXSTR], cbdecseq[MAXSTR]; int icosel, ni = getval("ni"), t1_counter; /* used for states tppi in t1 */ double tau1, /* t1 delay */ tau2, /* t2 delay */ tau3, /* t2 delay */ taua, /* ~ 1/4JNH = 2.25 ms */ taub, /* ~ 1/4JNH = 2.25 ms */ tauc, /* ~ 1/4JCaC' = 4 ms */ taud, /* ~ 1/4JCaC' = 4.5 ms if bigTCo can be set to be less than 4.5ms and then taud can be smaller*/ zeta, /* time for C'-N to refocuss set to 0.5*24.0 ms */ bigTCa, /* Ca T period */ bigTCo, /* Co T period */ bigTN, /* nitrogen T period */ BigT1, /* delay to compensate for gradient gt5 */ sw1, /* sweep width in f1 */ sw2, /* sweep width in f2 */ sphase, /* small angle phase shift */ sphase1, sphase2, /* used only for constant t2 period */ pwS4, /* selective CO 180 */ pwS3, /* selective Ca 180 */ pwS1, /* selecive Ca 90 */ pwS2, /* selective CO 90 */ cbpwr, /* power level for selective CB decoupling */ cbdmf, /* pulse width for selective CB decoupling */ cbres, /* decoupling resolution of CB decoupling */ gt1, gt2, gt3, gt4, gt5, gt6, gt7, gt8, gt9, gt10, gt11, gt12, gstab, gzlvl1, gzlvl2, gzlvl3, gzlvl4, gzlvl5, gzlvl6, gzlvl7, gzlvl8, gzlvl9, gzlvl10, gzlvl11, gzlvl12, compH = getval("compH"), /* adjustment for amplifier compression */ pwHs = getval ("pwHs"), /* H1 90 degree pulse at tpwrs */ tpwrs, /* power for pwHs ("H2osinc") pulse */ pwClvl = getval("pwClvl"), /* coarse power for C13 pulse */ pwC = getval("pwC"), /* C13 90 degree pulse length at pwClvl */ pwNlvl = getval("pwNlvl"), /* power for N15 pulses */ pwN = getval("pwN"), /* N15 90 degree pulse length at pwNlvl */ swCa = getval("swCa"), swCO = getval("swCO"), swN = getval("swN"), swTilt, /* This is the sweep width of the tilt vector */ cos_N, cos_CO, cos_Ca, angle_N, angle_CO, angle_Ca; angle_N=0.0; /*initialize variable*/ /* LOAD VARIABLES */ getstr("satmode",satmode); getstr("fc180",fc180); getstr("fscuba",fscuba); getstr("ddseq",ddseq); getstr("fCTCa",fCTCa); getstr("sel_flg",sel_flg); taua = getval("taua"); taub = getval("taub"); tauc = getval("tauc"); taud = getval("taud"); zeta = getval("zeta"); bigTCa = getval("bigTCa"); bigTCo = getval("bigTCo"); bigTN = getval("bigTN"); BigT1 = getval("BigT1"); tpwr = getval("tpwr"); dpwr = getval("dpwr"); dpwr3 = getval("dpwr3"); sw1 = getval("sw1"); sw2 = getval("sw2"); sphase = getval("sphase"); sphase1 = getval("sphase1"); sphase2 = getval("sphase2"); gt1 = getval("gt1"); gt2 = getval("gt2"); gt3 = getval("gt3"); gt4 = getval("gt4"); gt5 = getval("gt5"); gt6 = getval("gt6"); gt7 = getval("gt7"); gt8 = getval("gt8"); gt9 = getval("gt9"); gt10 = getval("gt10"); gt11 = getval("gt11"); gt12 = getval("gt12"); gstab = getval("gstab"); gzlvl1 = getval("gzlvl1"); gzlvl2 = getval("gzlvl2"); gzlvl3 = getval("gzlvl3"); gzlvl4 = getval("gzlvl4"); gzlvl5 = getval("gzlvl5"); gzlvl6 = getval("gzlvl6"); gzlvl7 = getval("gzlvl7"); gzlvl8 = getval("gzlvl8"); gzlvl9 = getval("gzlvl9"); gzlvl10 = getval("gzlvl10"); gzlvl11 = getval("gzlvl11"); gzlvl12 = getval("gzlvl12"); /* Load variable */ cbpwr = getval("cbpwr"); cbdmf = getval("cbdmf"); cbres = getval("cbres"); tau1 = 0; tau2 = 0; tau3 = 0; cos_N = 0; cos_CO = 0; cos_Ca = 0; getstr("cbdecseq", cbdecseq); /* LOAD PHASE TABLE */ settable(t1,1,phi1); settable(t2,1,phi2); settable(t3,4,phi3); settable(t4,1,phi4); settable(t5,1,phi5); settable(t7,4,phi7); settable(t8,4,phi8); settable(t6,4,rec); pwS1=c13pulsepw("ca", "co", "square", 90.0); pwS2=c13pulsepw("co", "ca", "sinc", 90.0); pwS3=c13pulsepw("ca", "co", "square", 180.0); pwS4=c13pulsepw("co", "ca", "sinc", 180.0); tpwrs = tpwr - 20.0*log10(pwHs/(compH*pw*1.69)); /*needs 1.69 times more*/ tpwrs = (int) (tpwrs); /*power than a square pulse */ /* CHECK VALIDITY OF PARAMETER RANGES */ if((dm[A] == 'y' || dm[B] == 'y' || dm[C] == 'y' )) { printf("incorrect dec1 decoupler flags! "); psg_abort(1); } if((dm2[A] == 'y' || dm2[B] == 'y' || dm2[C] == 'y')) { printf("incorrect dec2 decoupler flags! Should be 'nnn' "); psg_abort(1); } if( satpwr > 6 ) { printf("TSATPWR too large !!! "); psg_abort(1); } if( dpwr > 46 ) { printf("don't fry the probe, DPWR too large! "); psg_abort(1); } if( dpwr2 > 46 ) { printf("don't fry the probe, DPWR2 too large! "); psg_abort(1); } if( pwClvl > 62 ) { printf("don't fry the probe, pwClvl too large! "); psg_abort(1); } if( pw > 200.0e-6 ) { printf("dont fry the probe, pw too high ! "); psg_abort(1); } if( pwN > 200.0e-6 ) { printf("dont fry the probe, pwN too high ! "); psg_abort(1); } if( pwC > 200.0e-6 ) { printf("dont fry the probe, pwC too high ! "); psg_abort(1); } if( gt3 > 2.5e-3 ) { printf("gt3 is too long\n"); psg_abort(1); } if( gt1 > 10.0e-3 || gt2 > 10.0e-3 || gt4 > 10.0e-3 || gt5 > 10.0e-3 || gt6 > 10.0e-3 || gt7 > 10.0e-3 || gt8 > 10.0e-3 || gt9 > 10.0e-3 || gt10 > 10.0e-3 || gt11 > 50.0e-6) { printf("gt values are too long. Must be < 10.0e-3 or gt11=50us\n"); psg_abort(1); } /* PHASES AND INCREMENTED TIMES */ /* Set up angles and phases */ angle_CO=getval("angle_CO"); cos_CO=cos(PI*angle_CO/180.0); angle_Ca=getval("angle_Ca"); cos_Ca=cos(PI*angle_Ca/180.0); if ( (angle_CO < 0) || (angle_CO > 90) ) { printf ("angle_CO must be between 0 and 90 degree.\n"); psg_abort(1); } if ( (angle_Ca < 0) || (angle_Ca > 90) ) { printf ("angle_Ca must be between 0 and 90 degree.\n"); psg_abort(1); } if ( 1.0 < (cos_CO*cos_CO + cos_Ca*cos_Ca) ) { printf ("Impossible angles.\n"); psg_abort(1); } else { cos_N=sqrt(1.0- (cos_CO*cos_CO + cos_Ca*cos_Ca)); angle_N = 180.0*acos(cos_N)/PI; } swTilt=swCO*cos_CO + swCa*cos_Ca + swN*cos_N; if (ix ==1) { printf("\n\nn\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n"); printf ("Maximum Sweep Width: \t\t %f Hz\n", swTilt); printf ("Anlge_CO:\t%6.2f\n", angle_CO); printf ("Anlge_Ca:\t%6.2f\n", angle_Ca); printf ("Anlge_N :\t%6.2f\n", angle_N ); } /* Set up hyper complex */ /* sw1 is used as symbolic index */ if ( sw1 < 1000 ) { printf ("Please set sw1 to some value larger than 1000.\n"); psg_abort(1); } if (ix == 1) d2_init = d2; t1_counter = (int) ( (d2-d2_init)*sw1 + 0.5 ); if (t1_counter % 2) { tsadd(t2,2,4); tsadd(t6,2,4); } if (phase1 == 1) { ;} /* CC */ else if (phase1 == 2) { tsadd(t5,1,4);} /* SC */ else if (phase1 == 3) { tsadd(t1,1,4); } /* CS */ else if (phase1 == 4) { tsadd(t5,1,4); tsadd(t1,1,4); } /* SS */ else { printf ("phase1 can only be 1,2,3,4. \n"); psg_abort(1); } if (phase2 == 2) { tsadd(t4,2,4); icosel = 1; } /* N */ else icosel = -1; tau1 = 1.0*t1_counter*cos_Ca/swTilt; tau2 = 1.0*t1_counter*cos_CO/swTilt; tau3 = 1.0*t1_counter*cos_N/swTilt; tau1 = tau1/2.0; tau2 = tau2/2.0; tau3 = tau3/2.0; /* CHECK VALIDITY OF PARAMETER RANGES */ if (bigTN - 0.5*ni*(cos_N/swTilt) + pwS4 < 0.2e-6) { printf(" ni is too big. Make ni equal to %d or less.\n", ((int)((bigTN + pwS4)*2.0*swTilt/cos_N))); psg_abort(1);} if ((fCTCa[A]=='y') && (bigTCa - 0.5*ni*(cos_Ca/swTilt) - WFG_STOP_DELAY - POWER_DELAY - gt11 - 50.2e-6 < 0.2e-6)) { printf(" ni is too big for Ca. Make ni equal to %d or less.\n", (int) ((bigTCa -WFG_STOP_DELAY - POWER_DELAY - gt11 - 50.2e-6)/(0.5*cos_Ca/swTilt)) ); psg_abort(1); } if (bigTCo - 0.5*ni*(cos_CO/swTilt) - 4.0e-6 - POWER_DELAY < 0.2e-6) { printf(" ni is too big for CO. Make ni equal to %d or less.\n", (int) ((bigTCo - 4.0e-6 - POWER_DELAY) / (0.5*cos_CO/swTilt)) ); psg_abort(1); } /* BEGIN ACTUAL PULSE SEQUENCE */ status(A); obsoffset(tof); obspower(satpwr); /* Set transmitter power for 1H presaturation */ obspwrf(4095.0); decpower(pwClvl); /* Set Dec1 power for hard 13C pulses */ decpwrf(4095.0); dec2power(pwNlvl); /* Set Dec2 power for 15N hard pulses */ dec2pwrf(4095.0); set_c13offset("ca"); /* set Dec1 carrier at Ca */ sim3_c13pulse("", "ca", "co", "square", "", 0.0, 180.0, 0.0, zero, zero, zero, 2.0e-6, 0.0); set_c13offset("co"); /* set Dec1 carrier at Co */ /* Presaturation Period */ if (satmode[0] == 'y') { delay(2.0e-5); rgpulse(d1,zero,2.0e-6,2.0e-6); /* presaturation */ obspower(tpwr); /* Set transmitter power for hard 1H pulses */ delay(2.0e-5); if(fscuba[0] == 'y') { delay(2.2e-2); rgpulse(pw,zero,2.0e-6,0.0); rgpulse(2*pw,one,2.0e-6,0.0); rgpulse(pw,zero,2.0e-6,0.0); delay(2.2e-2); } } else { delay(d1); } obspower(tpwr); /* Set transmitter power for hard 1H pulses */ txphase(one); dec2phase(zero); delay(1.0e-5); /* Begin Pulses */ status(B); rcvroff(); lk_hold(); shiftedpulse("sinc", pwHs, 90.0, 0.0, one, 2.0e-6, 0.0); txphase(zero); delay(2.0e-6); /* xxxxxxxxxxxxxxxxxxxxxx 1HN to 15N TRANSFER xxxxxxxxxxxxxxxxxx */ rgpulse(pw,zero,0.0,0.0); /* 90 deg 1H pulse */ delay(0.2e-6); zgradpulse(gzlvl1, gt1); delay(2.0e-6); delay(taua - gt1 - 2.2e-6); /* taua <= 1/4JNH */ sim3pulse(2*pw,0.0,2*pwN,zero,zero,zero,0.0,0.0); txphase(three); dec2phase(zero); decphase(zero); delay(taua - gt1 - gstab -0.2e-6 - 2.0e-6); delay(0.2e-6); zgradpulse(gzlvl1, gt1); delay(gstab); /* xxxxxxxxxxxxxxxxxxxxxx 15N to 13CO TRANSFER xxxxxxxxxxxxxxxxxx */ if(sel_flg[A] == 'n') { rgpulse(pw,three,2.0e-6,0.0); delay(0.2e-6); zgradpulse(gzlvl2, gt2); delay(gstab); dec2rgpulse(pwN,zero,0.0,0.0); delay( zeta + pwS4 ); dec2rgpulse(2*pwN,zero,0.0,0.0); c13pulse("co", "ca", "sinc", 180.0, zero, 0.0, 0.0); dec2phase(one); delay(zeta - 2.0e-6); dec2rgpulse(pwN,one,2.0e-6,0.0); } else { rgpulse(pw,one,2.0e-6,0.0); initval(1.0,v6); dec2stepsize(45.0); dcplr2phase(v6); delay(0.2e-6); zgradpulse(gzlvl2, gt2); delay(gstab); dec2rgpulse(pwN,zero,0.0,0.0); dcplr2phase(zero); dec2phase(zero); delay(1.34e-3 - SAPS_DELAY - 2.0*pw); rgpulse(pw,one,0.0,0.0); rgpulse(2.0*pw,zero,0.0,0.0); rgpulse(pw,one,0.0,0.0); delay( zeta - 1.34e-3 - 2.0*pw + pwS4 ); dec2rgpulse(2*pwN,zero,0.0,0.0); c13pulse("co", "ca", "sinc", 180.0, zero, 0.0, 0.0); dec2phase(one); delay(zeta - 2.0e-6); dec2rgpulse(pwN,one,2.0e-6,0.0); } dec2phase(zero); decphase(zero); delay(0.2e-6); zgradpulse(gzlvl3, gt3); delay(gstab); /* xxxxxxxxxxxxxxxxxxxxx 13CO to 13CA TRANSFER xxxxxxxxxxxxxxxxxxxxxxx */ c13pulse("co", "ca", "sinc", 90.0, zero, 2.0e-6, 0.0); delay(2.0e-7); zgradpulse(gzlvl10, gt10); delay(100.0e-6); delay(tauc - POWER_DELAY - gt10 - 100.2e-6 - (0.5*10.933*pwC)); decrgpulse(pwC*158.0/90.0, zero, 0.0, 0.0); decrgpulse(pwC*171.2/90.0, two, 0.0, 0.0); decrgpulse(pwC*342.8/90.0, zero, 0.0, 0.0); /* Shaka 6 composite */ decrgpulse(pwC*145.5/90.0, two, 0.0, 0.0); decrgpulse(pwC*81.2/90.0, zero, 0.0, 0.0); decrgpulse(pwC*85.3/90.0, two, 0.0, 0.0); delay(2.0e-7); zgradpulse(gzlvl10, gt10); delay(100.0e-6); delay(tauc - POWER_DELAY - 4.0e-6 - gt10 - 100.2e-6 - (0.5*10.933*pwC)); c13pulse("co", "ca", "sinc", 90.0, one, 4.0e-6, 0.0); set_c13offset("ca"); /* change Dec1 carrier to Ca (55 ppm) */ delay(0.2e-6); zgradpulse(gzlvl9, gt9); delay(gstab); /* xxxxxxxxxxxxxxxxxx 13CA EVOLUTION xxxxxxxxxxxxxxxxxxxxxx */ /* Turn on D decoupling using the third decoupler */ dec3unblank(); dec3rgpulse(1/dmf3, one, 0.0, 0.0); dec3unblank(); setstatus(DEC3ch, TRUE, 'w', FALSE, dmf3); /* Turn on D decoupling */ c13pulse("ca", "co", "square", 90.0, t5, 2.0e-6, 0.0); if (fCTCa[A]=='y') { /* Constant t2 */ decpower(cbpwr); decphase(zero); decprgon(cbdecseq,1/cbdmf,cbres); decon(); delay(tau1); decoff(); decprgoff(); decpower(pwClvl); dec2rgpulse(pwN,one,0.0,0.0); dec2rgpulse(2*pwN,zero,0.0,0.0); dec2rgpulse(pwN,one,0.0,0.0); c13pulse("co", "ca", "sinc", 180.0, zero, 0.0, 0.0); decpower(cbpwr); decphase(zero); decprgon(cbdecseq,1/cbdmf,cbres); decon(); delay(bigTCa - 4.0*pwN - WFG_START_DELAY - pwS4 - WFG_STOP_DELAY - POWER_DELAY - WFG_START_DELAY - gt11 - gstab -0.2e-6); decoff(); decprgoff(); decpower(pwClvl); delay(0.2e-6); zgradpulse(gzlvl11, gt11); delay(gstab); initval(1.0,v3); decstepsize(140); dcplrphase(v3); c13pulse("ca", "co", "square", 180.0, zero, 0.0, 0.0); delay(0.2e-6); zgradpulse(gzlvl11, gt11); delay(gstab); decpower(cbpwr); decphase(zero); decprgon(cbdecseq,1/cbdmf,cbres); decon(); delay(bigTCa - tau1 - WFG_STOP_DELAY - POWER_DELAY - gt11 - gstab -0.2e-6); decoff(); decprgoff(); } /* non_constant t2 */ else { if (fc180[A]=='n') { if ((ni>1.0) && (tau1>0.0)) { if (tau1 - 2.0*pwS1/PI - PRG_START_DELAY - 2*POWER_DELAY - PRG_STOP_DELAY - pwN > 0.0) { decpower(cbpwr); decphase(zero); decprgon(cbdecseq,1/cbdmf,cbres); decon(); delay(tau1 - 2.0*pwS1/PI - PRG_START_DELAY - 2*POWER_DELAY - PRG_STOP_DELAY - pwN); decoff(); decprgoff(); decphase(zero); dec2phase(zero); decpower(pwClvl); sim3_c13pulse("", "co", "ca", "sinc", "", 0.0, 180.0, 2.0*pwN, zero, zero, zero, 0.0, 0.0); decpower(cbpwr); decphase(zero); decprgon(cbdecseq,1/cbdmf,cbres); decon(); delay(tau1 - 2.0*pwS1/PI - PRG_START_DELAY - 2*POWER_DELAY - PRG_STOP_DELAY - pwN); decoff(); decprgoff(); decstepsize(1.0); initval(sphase1, v3); dcplrphase(v3); } else { tsadd(t6,2,4); delay(2.0*tau1); delay(10.0e-6); sim3_c13pulse("", "ca", "co", "square", "", 0.0, 180.0, 2.0*pwN, zero, zero, zero, 2.0e-6, 0.0); delay(10.0e-6); } } else { tsadd(t6,2,4); delay(10.0e-6); sim3_c13pulse("", "ca", "co", "square", "", 0.0, 180.0, 2.0*pwN, zero, zero, zero, 2.0e-6, 0.0); delay(10.0e-6); } } else { /* for checking sequence */ c13pulse("ca", "co", "square", 180.0, zero, 2.0e-6, 0.0); } } decpower(pwClvl); decphase(t7); c13pulse("ca", "co", "square", 90.0, t7, 4.0e-6, 0.0); dcplrphase(zero); /* Turn off D decoupling */ dec3rgpulse(1/dmf3, three, 0.0, 0.0); dec3blank(); setstatus(DEC3ch, FALSE, 'w', FALSE, dmf3); dec3blank(); /* Turn off D decoupling */ set_c13offset("co"); /* set carrier back to Co */ delay(0.2e-6); zgradpulse(gzlvl12, gt12); delay(gstab); /* xxxxxxxxxxxxxxx 13CA to 13CO TRANSFER and CT 13CO EVOLUTION xxxxxxxxxxxxxxxxx */ c13pulse("co", "ca", "sinc", 90.0, t1, 2.0e-6, 0.0); delay(tau2); dec2rgpulse(pwN,one,0.0,0.0); dec2rgpulse(2*pwN,zero,0.0,0.0); dec2rgpulse(pwN,one,0.0,0.0); delay(taud - 4.0*pwN - POWER_DELAY - 0.5*(WFG_START_DELAY + pwS3 + WFG_STOP_DELAY)); c13pulse("ca", "co", "square", 180.0, zero, 0.0, 0.0); decphase(t8); initval(1.0,v4); decstepsize(sphase); dcplrphase(v4); delay(bigTCo - taud - 0.5*(WFG_START_DELAY + pwS3 + WFG_STOP_DELAY) ); c13pulse("co", "ca", "sinc", 180.0, t8, 0.0, 0.0); dcplrphase(zero); decphase(one); delay(bigTCo - tau2 - POWER_DELAY - 4.0e-6); c13pulse("co", "ca", "sinc", 90.0, one, 4.0e-6, 0.0); delay(0.2e-6); zgradpulse(gzlvl4, gt4); delay(gstab); /* t3 period */ dec2rgpulse(pwN,t2,2.0e-6,0.0); dec2phase(t3); delay(bigTN - tau3 + pwS4); dec2rgpulse(2*pwN,t3,0.0,0.0); c13pulse("co", "ca", "sinc", 180.0, zero, 0.0, 0.0); txphase(zero); dec2phase(t4); delay(bigTN - gt5 - gstab -0.2e-6 - 2.0*GRADIENT_DELAY - 4.0e-6 - WFG_START_DELAY - pwS3 - WFG_STOP_DELAY); delay(0.2e-6); zgradpulse(icosel*gzlvl5, gt5); delay(gstab); c13pulse("ca", "co", "square", 180.0, zero, 4.0e-6, 0.0); delay(tau3); sim3pulse(pw,0.0,pwN,zero,zero,t4,0.0,0.0); delay(0.2e-6); zgradpulse(gzlvl6, gt6); delay(2.0e-6); dec2phase(zero); delay(taub - gt6 - 2.2e-6); sim3pulse(2*pw,0.0,2*pwN,zero,zero,zero,0.0,0.0); delay(0.2e-6); zgradpulse(gzlvl6, gt6); delay(200.0e-6); txphase(one); dec2phase(one); delay(taub - gt6 - 200.2e-6); sim3pulse(pw,0.0,pwN,one,zero,one,0.0,0.0); delay(0.2e-6); zgradpulse(gzlvl7, gt7); delay(2.0e-6); txphase(zero); dec2phase(zero); delay(taub - gt7 - 2.2e-6); sim3pulse(2*pw,0.0,2*pwN,zero,zero,zero,0.0,0.0); delay(0.2e-6); zgradpulse(gzlvl7, gt7); delay(200.0e-6); delay(taub - gt7 - 200.2e-6); sim3pulse(pw,0.0,pwN,zero,zero,zero,0.0,0.0); delay(0.2e-6); zgradpulse(-gzlvl8, gt8/2.0); delay(50.0e-6); delay(BigT1 - gt8/2.0 - 50.2e-6 - 0.5*(pwN - pw) - 2.0*pw/PI); rgpulse(2*pw,zero,0.0,0.0); delay(0.2e-6); zgradpulse(gzlvl8, gt8/2.0); delay(50.0e-6); dec2power(dpwr2); decpower(dpwr); delay(BigT1 - gt8/2.0 - 50.2e-6 - 2.0*POWER_DELAY); lk_sample(); /* rcvron(); */ /* Turn on receiver to warm up before acq */ /* BEGIN ACQUISITION */ status(C); setreceiver(t6); }

pulsesequence() { // Define Variables and Objects and Get Parameter Values CP hy = getcp("HY",0.0,0.0,0,1); strncpy(hy.fr,"dec",3); strncpy(hy.to,"dec2",4); putCmd("frHY='dec'\n"); putCmd("toHY='dec2'\n"); GP inept = getinept("ineptYX"); strncpy(inept.ch1,"dec2",4); strncpy(inept.ch2,"obs",3); putCmd("ch1YXinept='dec2'\n"); putCmd("ch2YXinept='obs'\n"); DSEQ dec = getdseq("H"); strncpy(dec.t.ch,"dec",3); putCmd("chHtppm='dec'\n"); strncpy(dec.s.ch,"dec",3); putCmd("chHspinal='dec'\n"); DSEQ mix = getdseq("Hmix"); strncpy(mix.t.ch,"dec",3); putCmd("chHmixtppm='dec'\n"); strncpy(mix.s.ch,"dec",3); putCmd("chHmixspinal='dec'\n"); // Dutycycle Protection double simpw1 = inept.pw1; if (inept.pw2 > inept.pw1) simpw1 = inept.pw2; double simpw2 = inept.pw3; if (inept.pw4 > inept.pw3) simpw2 = inept.pw4; DUTY d = init_dutycycle(); d.dutyon = getval("pwH90") + getval("tHY") + 2.0*simpw1 + 2.0*simpw2; d.dutyoff = d1 + 4.0e-6; d.c1 = d.c1 + (!strcmp(dec.seq,"tppm")); d.c1 = d.c1 + ((!strcmp(dec.seq,"tppm")) && (dec.t.a > 0.0)); d.t1 = inept.t1 + inept.t2 + inept.t3 + inept.t4 + getval("rd") + getval("ad") + at; d.c2 = d.c2 + (!strcmp(dec.seq,"spinal")); d.c2 = d.c2 + ((!strcmp(dec.seq,"spinal")) && (dec.s.a > 0.0)); d.t2 = inept.t1 + inept.t2 + inept.t3 + inept.t4 + getval("rd") + getval("ad") + at; d = update_dutycycle(d); abort_dutycycle(d,10.0); // Set Phase Tables settable(phH90,16,table1); settable(phHhy,4,table2); settable(phYhy,4,table3); settable(ph1Yyxinept,4,table4); settable(ph1Xyxinept,4,table5); settable(ph2Yyxinept,4,table6); settable(ph2Xyxinept,16,table7); settable(ph3Yyxinept,8,table8); settable(ph3Xyxinept,4,table9); settable(phRec,8,table10); setreceiver(phRec); // Begin Sequence txphase(ph1Xyxinept); decphase(phH90); dec2phase(phYhy); obspwrf(getval("aXyxinept")); decpwrf(getval("aH90")); dec2pwrf(getval("aYhy")); obsunblank(); decunblank(); _unblank34(); delay(d1); sp1on(); delay(2.0e-6); sp1off(); delay(2.0e-6); // H to Y Cross Polarization decrgpulse(getval("pwH90"),phH90,0.0,0.0); decphase(phHhy); _cp_(hy,phHhy,phYhy); decphase(zero); // INEPT Transfer from Y to X _dseqon(mix); _ineptref(inept,ph1Yyxinept,ph1Xyxinept,ph2Yyxinept,ph2Xyxinept,ph3Yyxinept,ph3Xyxinept); _dseqoff(mix); // Begin Acquisition _dseqon(dec); obsblank(); _blank34(); delay(getval("rd")); startacq(getval("ad")); acquire(np, 1/sw); endacq(); _dseqoff(dec); obsunblank(); decunblank(); _unblank34(); }

pulsesequence() { // Set the Maximum Dynamic Table and v-var Numbers settablenumber(10); setvvarnumber(30); // Define Variables and Objects and Get Parameter Values double aXprep1 = getval("aXprep1"); // Define Tilted Pulses using "prep1X". double pw1Xprep1 = getval("pw1Xprep1"); double pw2Xprep1 = getval("pw2Xprep1"); double phXprep1 = getval("phXprep1"); WMPA wpmlg = getwpmlg("wpmlgX"); strncpy(wpmlg.ch,"obs",3); putCmd("chXwpmlg='obs'\n"); //-------------------------------------- // Copy Current Parameters to Processed //------------------------------------- putCmd("groupcopy('current','processed','acquisition')"); // Dutycycle Protection DUTY d = init_dutycycle(); d.dutyon = getval("pw1Xprep1") + getval("pw2Xprep1") + 2.0*wpmlg.q*wpmlg.cycles*wpmlg.pw; d.dutyoff = d1 + 4.0e-6 + 5.0e-6 + wpmlg.r1 + wpmlg.r2 + at - 2.0*wpmlg.q*wpmlg.cycles*wpmlg.pw; d = update_dutycycle(d); abort_dutycycle(d,10.0); // Set Phase Tables settable(ph1Xprep1,4,table1); settable(ph2Xprep1,4,table2); settable(phXwpmlg,4,table3); settable(phRec,4,table4); setreceiver(phRec); // Set the Small-Angle Step double obsstep = 360.0/(PSD*8192); obsstepsize(obsstep); int phfXprep1 = initphase(phXprep1, obsstep); int phXzero = initphase(0.0, obsstep); // Begin Sequence xmtrphase(phfXprep1); txphase(ph1Xprep1); obspwrf(aXprep1); obsunblank(); decunblank(); _unblank34(); delay(d1); sp1on(); delay(2.0e-6); sp1off(); delay(2.0e-6); // Tilted Preparation Pulse for FSLG or PMLG "prep1X" startacq(5.0e-6); rcvroff(); delay(wpmlg.r1); rgpulse(pw1Xprep1, ph1Xprep1, 0.0, 0.0); rgpulse(pw2Xprep1, ph2Xprep1, 0.0, 0.0); xmtrphase(phXzero); delay(wpmlg.r2); // Apply WPMLG Cycles decblank(); _blank34(); _wpmlg(wpmlg, phXwpmlg); endacq(); obsunblank(); decunblank(); _unblank34(); }