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view rDiff/src/tools/read_utils/get_reads_direct.cpp @ 0:0f80a5141704
version 0.3 uploaded
author | vipints |
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date | Thu, 14 Feb 2013 23:38:36 -0500 |
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/* written by Jonas Behr, Regina Bohnert and Gunnar Raetsch, FML Tuebingen, Germany, 2010 */ #include <stdio.h> #include <assert.h> #include "sam.h" #include "get_reads_direct.h" #include <vector> using std::vector; #include <string> using std::string; #ifdef __cplusplus extern "C" { #endif typedef struct { int beg, end; samfile_t *in; } tmpstruct_t; typedef struct { uint64_t u, v; } pair64_t; static inline int is_overlap(uint32_t beg, uint32_t end, const bam1_t *b) { uint32_t rbeg = b->core.pos; uint32_t rend = b->core.n_cigar? bam_calend(&b->core, bam1_cigar(b)) : b->core.pos + 1; return (rend > beg && rbeg < end); } pair64_t * get_chunk_coordinates(const bam_index_t *idx, int tid, int beg, int end, int* cnt_off); int bam_fetch_reads(bamFile fp, const bam_index_t *idx, int tid, int beg, int end, void *data, bam_header_t* header, vector<CRead*>* reads, char strand); // callback for bam_plbuf_init() static int pileup_func(uint32_t tid, uint32_t pos, int n, const bam_pileup1_t *pl, void *data) { //tmpstruct_t *tmp = (tmpstruct_t*)data; //if ((int)pos >= tmp->beg && (int)pos < tmp->end) // printf("%s\t%d\t%d\n", tmp->in->header->target_name[tid], pos + 1, n); return 0; } #ifdef __cplusplus } #endif int parse_sam_line(char* line, CRead* read); //int set_strand(char c); //void parse_cigar(bam1_t* b, CRead* read); int get_reads_from_bam(char* filename, char* region, vector<CRead*>* reads, char strand, int lsubsample) { subsample = lsubsample; //set_strand(strand); srand (time(NULL)); //srand (1234); tmpstruct_t tmp; tmp.in = samopen(filename, "rb", 0); if (tmp.in == 0) { fprintf(stderr, "Fail to open BAM file %s\n", filename); return 1; } int ref; bam_index_t *idx; bam_plbuf_t *buf; idx = bam_index_load(filename); // load BAM index if (idx == 0) { fprintf(stderr, "BAM indexing file is not available.\n"); samclose(tmp.in); return 1; } bam_parse_region(tmp.in->header, region, &ref, &tmp.beg, &tmp.end); // parse the region if (ref < 0) { fprintf(stderr, "Invalid region %s\n", region); bam_index_destroy(idx); samclose(tmp.in); return 1; } buf = bam_plbuf_init(pileup_func, &tmp); // initialize pileup bam_fetch_reads(tmp.in->x.bam, idx, ref, tmp.beg, tmp.end, buf, tmp.in->header, reads, strand); //fprintf(stdout, "intron_list: %d \n", intron_list->size()); bam_plbuf_push(0, buf); // finalize pileup bam_index_destroy(idx); bam_plbuf_destroy(buf); samclose(tmp.in); return 0; } int bam_fetch_reads(bamFile fp, const bam_index_t *idx, int tid, int beg, int end, void *data, bam_header_t* header, vector<CRead*>* reads, char strand) { int n_off; pair64_t *off = get_chunk_coordinates(idx, tid, beg, end, &n_off); if (off == 0) return 0; { // retrive alignments uint64_t curr_off; int i, ret, n_seeks; n_seeks = 0; i = -1; curr_off = 0; bam1_t *b = (bam1_t*)calloc(1, sizeof(bam1_t)); for (;;) { if (curr_off == 0 || curr_off >= off[i].v) { // then jump to the next chunk if (i == n_off - 1) break; // no more chunks if (i >= 0) assert(curr_off == off[i].v); // otherwise bug if (i < 0 || off[i].v != off[i+1].u) { // not adjacent chunks; then seek bam_seek(fp, off[i+1].u, SEEK_SET); curr_off = bam_tell(fp); ++n_seeks; } ++i; } if ((ret = bam_read1(fp, b)) > 0) { curr_off = bam_tell(fp); if (b->core.tid != tid || b->core.pos >= end) break; // no need to proceed else if (is_overlap(beg, end, b)) { int rr = rand(); if ((rr%1000 < subsample)) { CRead* read = new CRead(); parse_cigar(b, read, header); if (strand == '0' || strand==read->strand[0] || read->strand[0]=='0') { reads->push_back(read); } else { delete read; } //else if (read->strand[0]=='0'&&((b->core.flag & g_flag_off) >0)) //{ // //fprintf(stdout, "(-)-strand; read->strand[0]==0, num_exons: %i \n", read->block_starts.size()); // // this flag means that the read has been reversed for alignment // // flag bit set and (-)-strand requested // reads->push_back(read); //} //else if (read->strand[0]=='0'&&(g_flag_on>0&&(b->core.flag & g_flag_on)==0)) //{ // //fprintf(stdout, "(+)-strand; read->strand[0]==0, num_exons: %i \n", read->block_starts.size()); // // (+)-strand requested and flag bit not set // reads->push_back(read); //} } } } else break; // end of file } // fprintf(stderr, "[bam_fetch] # seek calls: %d\n", n_seeks); bam_destroy1(b); } free(off); return 0; } void parse_cigar(bam1_t* b, CRead* read, bam_header_t* header) { read->left = (b->core.flag & left_flag_mask) >0; read->right = (b->core.flag & right_flag_mask) >0; read->reverse = (b->core.flag & reverse_flag_mask) >0; read->start_pos = b->core.pos+1; read->set_strand('0'); read->read_id = new char[1000]; //sprintf(read->read_id, "%s\0", bam1_qname(b)); sprintf(read->read_id, "%s", bam1_qname(b)); for (int k = 0; k < b->core.n_cigar; ++k) { int op = bam1_cigar(b)[k] & BAM_CIGAR_MASK; // operation int l = bam1_cigar(b)[k] >> BAM_CIGAR_SHIFT; // length //int op = bam_cigar_op(bam1_cigar(b)[k]); // operation //int l = bam_cigar_oplen(bam1_cigar(b)[k]); // length //fprintf(stdout, "op:%d l:%d\n", op, l); if (op == BAM_CMATCH) { if (k==0) { read->block_lengths.push_back(l); read->block_starts.push_back(0); } else { int op_prev = bam1_cigar(b)[k-1] & BAM_CIGAR_MASK; int l_prev = bam1_cigar(b)[k-1] >> BAM_CIGAR_SHIFT; if (op_prev==BAM_CREF_SKIP)// intron before { if (read->block_lengths.size()>=1) { int last_block_start = (*(read->block_starts.end()-1)); int intron_start = last_block_start+(*(read->block_lengths.end()-1)); read->block_lengths.push_back(l); read->block_starts.push_back(intron_start+l_prev); } else { // start of first block was not a match read->block_lengths.push_back(l); read->block_starts.push_back(0); } } else { if (read->block_lengths.size()>=1) (*(read->block_lengths.end()-1))+=l; else { read->block_lengths.push_back(l); read->block_starts.push_back(0); } } } } else if (op == BAM_CDEL) { if (k>0 && read->block_lengths.size()>=1) (*(read->block_lengths.end()-1))+=l; } else if (op == BAM_CREF_SKIP)//intron {} else if (op == BAM_CINS) {} else if (op == BAM_CSOFT_CLIP || op == BAM_CHARD_CLIP) { read->is_clipped = true; } } // parse auxiliary data uint8_t* s = bam1_aux(b); uint8_t* end = b->data + b->data_len; while (s < end) { uint8_t type, key[2]; key[0] = s[0]; key[1] = s[1]; s += 2; type = *s; ++s; //fprintf(stdout, "\n%c%c:%c\n", key[0], key[1], type); if (type == 'A') { if ( key[0] =='X' && key[1] == 'S') { read->set_strand((char) *s); } ++s; } else if (type == 'C') { if ( key[0] =='H' && key[1] == '0') { uint8_t matches = *s; read->matches = (int) matches; } if ( key[0] =='N' && key[1] == 'M') { uint8_t mismatches = *s; read->mismatches = (int) mismatches; } if ( key[0] =='H' && key[1] == 'I') { uint8_t mai = *s; read->multiple_alignment_index = (int) mai; } ++s; } else if (type == 'c') { ++s; } else if (type == 'S') { s += 2; } else if (type == 's') { s += 2; } else if (type == 'I') { s += 4; } else if (type == 'i') { s += 4; } else if (type == 'f') { s += 4; } else if (type == 'd') { s += 8; } else if (type == 'Z') { ++s; } else if (type == 'H') { ++s; } } } //int set_strand(char c) //{ // if (c=='+') // { // char* fl = (char*) "0x0010"; // g_flag_on = strtol(fl, 0, 0); // g_flag_off = 0; // } // else if (c=='-') // { // char* fl = (char*) "0x0010"; // g_flag_off = strtol(fl, 0, 0); // g_flag_on = 0; // } // return 0; //}