sff_t *
sff_read1(FILE *fp, sff_header_t *header)
{
sff_t *sff;
sff = sff_init();
sff->gheader = header;
sff->rheader = sff_read_header_read(fp);
if(NULL == sff->rheader) { // EOF
sff_destroy(sff);
return NULL;
}
sff->read = sff_read_read(fp, sff->gheader, sff->rheader);
if(NULL == sff->read) { // EOF
sff_destroy(sff);
return NULL;
}
return sff;
}
int main(int argc, char *argv[])
{
char *fastqFileName = NULL;
char *sffFileName = NULL;
bool forceClip = false;
bool keyPass = false;
bool allReads = false;
int readCol = -1;
int readRow = -1;
bool findRead = false;
int row, col;
int numKeypassedReads = 0;
int qual_offset = DEFAULT_QUAL_OFFSET;
bool legacyFASTQName = false; // enable if you want r10|c100 format name in fastq file
bool debug = false;
bool legacyReadName = false;
bool adapterTrim = true;
bool ignoreLeftQualTrim = false;
// process command-line args
int argcc = 1;
while (argcc < argc) {
if (argv[argcc][0] == '-') {
switch (argv[argcc][1]) {
case 'a': // output all reads
allReads = true;
break;
case 'R': // report read at row & column
argcc++;
readRow = atoi(argv[argcc]);
break;
case 'C': // report read at row & column
argcc++;
readCol = atoi(argv[argcc]);
break;
case 'q': // convert to fastq
argcc++;
fastqFileName = argv[argcc];
break;
case 'c': // force qual clip left to 5
forceClip = true;
break;
case 's': // Offset to apply to quality scores
argcc++;
qual_offset = atoi(argv[argcc]);
if(qual_offset==0) {
fprintf (stderr, "-s option should specify a nonzero quality offset\n");
exit (1);
}
break;
case 'k': // force keypass
keyPass = true;
argcc++;
hackkey = argv[argcc];
hackkeylen = strlen(hackkey);
break;
case 'L': // don't record name of read in comment
legacyFASTQName = true;
break;
case 'd': // enable debug print outs
debug = true;
break;
case 'h': // help info
printHelp ();
exit (0);
break;
case 'u': // prevent read clipping
adapterTrim = false;
break;
case 'b': // ignore barcodes (ok really its ignoring the left qual trim)
ignoreLeftQualTrim = true;
break;
case 'v': // version info
fprintf (stdout, "%s", IonVersion::GetFullVersion("SFFRead").c_str());
exit (0);
break;
default:
//sffFileName = argv[argcc];
break;
}
}
else {
sffFileName = argv[argcc];
}
argcc++;
}
if (!sffFileName) {
printHelp();
exit(0);
}
if (readCol > -1 && readRow > -1) {
findRead = true;
allReads = true;// makes it search all reads
}
sff_file_t* sff_file_in = NULL;
sff_file_in = sff_fopen(sffFileName, "rb", NULL, NULL);
if (sff_file_in) {
if (!findRead && !fastqFileName) {
printf("Reading file: %s\n", sffFileName);
sff_header_print(stdout, sff_file_in->header);
}
// -- read the reads
int numReads;
if (allReads) {
numReads = sff_file_in->header->n_reads;
}
else {
numReads = (sff_file_in->header->n_reads < 10 ? sff_file_in->header->n_reads:10);
}
FILE *fpq = NULL;
if (fastqFileName) {
numReads = sff_file_in->header->n_reads;
fpq = fopen(fastqFileName, "w");
if (!fpq){
perror (fastqFileName);
exit (1);
}
}
for(int i=0;i<numReads;i++) {
sff_read_header_t* rh = sff_read_header_read(sff_file_in->fp);
sff_read_t* rr = sff_read_read(sff_file_in->fp, sff_file_in->header, rh);
// optional - ignore the left & right adapter clipping by simply setting these values to 0
if (!adapterTrim) {
rh->clip_adapter_left = 0;
rh->clip_adapter_right = 0;
}
if (!fpq && !findRead) {
printf("Read header length: %d\n", rh->rheader_length);
printf("Read name length: %d\n", rh->name_length);
}
// Extract the row and column popsition info for this read
if (1 != ion_readname_to_rowcol(rh->name->s, &row, &col)) {
fprintf (stderr, "Error parsing read name: '%s'\n", rh->name->s);
continue;
}
if(1 == ion_readname_legacy(rh->name->s)) {
legacyReadName = true;
}
else {
legacyReadName = false;
}
if (!fpq && !findRead) {
printf("Read: %s (r%05d|c%05d) has %d bases\n",
(rh->name_length > 0 ? rh->name->s : "NONAME"),
row, col,
rh->n_bases);
printf("Clip left: %d qual: %d right: %d qual: %d\n",
rh->clip_adapter_left, rh->clip_qual_left,
rh->clip_adapter_right, rh->clip_qual_right);
printf("Flowgram values:\n");
}
if (findRead) {
if (row == readRow && col == readCol) {
//printf("Ionogram: ");
int i;
for(i=0;i<sff_file_in->header->flow_length;i++) {
printf("%.2lf ", (double)(rr->flowgram[i])/100.0);
}
printf("\n");
//// now print the bases - all the bases, not clipped!
//// these bases correspond to the raw flowgram data. in essence
//for (int b=0;b<r.number_of_bases;b++)
// fprintf(stdout, "%c", bases[b]);
//fprintf(stdout, "\n");
}
}
else if (fpq) {
bool ok = true;
if (keyPass) {
// if (r.number_of_bases > h.key_length) {
if ((int)rh->n_bases > hackkeylen) {
int b;
// for(b=0;b<h.key_length;b++) {
for(b=0;b<hackkeylen;b++) {
// if (key_sequence[b] != bases[b]) {
if (hackkey[b] != rr->bases->s[b]) {
ok = false;
break;
}
}
} else
ok = false; // not long enough
}
int clip_left_index = 0;
int clip_right_index = 0;
if (ok) {
//numKeypassedReads++;
// If force-clip option is set, we want to ensure the key gets trimmed
if (forceClip && rh->clip_adapter_left < 4)
rh->clip_adapter_left = hackkeylen+1;
if (ignoreLeftQualTrim)
clip_left_index = max (1, rh->clip_adapter_left);
else
clip_left_index = max (1, max (rh->clip_qual_left, rh->clip_adapter_left));
clip_right_index = min ((rh->clip_qual_right == 0 ? rh->n_bases:rh->clip_qual_right),
(rh->clip_adapter_right == 0 ? rh->n_bases:rh->clip_adapter_right));
if (debug)
fprintf (stdout, "debug clip: left = %d right = %d\n", clip_left_index, clip_right_index);
numKeypassedReads++;
if (clip_left_index > clip_right_index)
// Suppress output of zero-mer reads (left > right)
ok = false;
}
if (ok) {
//print id string
if (legacyFASTQName) {
fprintf (fpq, "@r%d|c%d\n", row, col);
}
else {
if (legacyReadName){
//Override legacy name
char runId[6] = {'\0'};
strncpy (runId, &rh->name->s[7], 5);
fprintf (fpq, "@%s:%d:%d\n", runId, row, col);
}
else {
//Copy name verbatim
fprintf (fpq, "@%s\n", rh->name->s);
}
}
//print bases
for (int b=clip_left_index-1;b<clip_right_index;b++)
fprintf(fpq, "%c", rr->bases->s[b]);
fprintf(fpq, "\n");
//print '+'
fprintf(fpq, "+\n");
//print quality scores
for (int b=clip_left_index-1;b<clip_right_index;b++)
fprintf(fpq, "%c", QualToFastQ((int)(rr->quality->s[b]),qual_offset));
fprintf(fpq, "\n");
}
}
else {
int f;
for(f=0;f<sff_file_in->header->flow_length;f++)
printf("%d ", rr->flowgram[f]);
printf("\nFlow index per base:\n");
unsigned int b;
for(b=0;b<rh->n_bases;b++)
printf("%d ", rr->flow_index[b]);
printf("\nBases called:\n");
for(b=0;b<rh->n_bases;b++)
printf("%c", rr->bases->s[b]);
printf("\nQuality scores:\n");
for(b=0;b<rh->n_bases;b++)
printf("%d ", rr->quality->s[b]);
printf("\nDone with this read\n\n");
}
sff_read_header_destroy(rh);
sff_read_destroy(rr);
}
// debug print - keypass reads written to the fastq file
if (fpq) {
static char *printkey = "All";
if (keyPass) printkey = hackkey;
fprintf (stdout, "Keypass Reads(%s) = %d\n", printkey, numKeypassedReads);
fprintf (stdout, "Total Reads = %d\n", numReads);
fprintf (stdout, "Percentage = %.2f%%\n", ((float) numKeypassedReads/ (float) numReads) * 100.0);
}
sff_fclose(sff_file_in);
if (fpq)
fclose(fpq);
}
else {
perror (sffFileName);
exit (1);
}
return 0;
}
