| 0 | 1 #include <unistd.h> | 
|  | 2 #include <assert.h> | 
|  | 3 #include <string.h> | 
|  | 4 #include <ctype.h> | 
|  | 5 #include <math.h> | 
|  | 6 #include "faidx.h" | 
|  | 7 #include "sam.h" | 
|  | 8 #include "kstring.h" | 
|  | 9 #include "kaln.h" | 
|  | 10 #include "kprobaln.h" | 
|  | 11 | 
|  | 12 #define USE_EQUAL 1 | 
|  | 13 #define DROP_TAG  2 | 
|  | 14 #define BIN_QUAL  4 | 
|  | 15 #define UPDATE_NM 8 | 
|  | 16 #define UPDATE_MD 16 | 
|  | 17 #define HASH_QNM  32 | 
|  | 18 | 
|  | 19 char bam_nt16_nt4_table[] = { 4, 0, 1, 4, 2, 4, 4, 4, 3, 4, 4, 4, 4, 4, 4, 4 }; | 
|  | 20 | 
|  | 21 int bam_aux_drop_other(bam1_t *b, uint8_t *s); | 
|  | 22 | 
|  | 23 void bam_fillmd1_core(bam1_t *b, char *ref, int flag, int max_nm) | 
|  | 24 { | 
|  | 25 	uint8_t *seq = bam1_seq(b); | 
|  | 26 	uint32_t *cigar = bam1_cigar(b); | 
|  | 27 	bam1_core_t *c = &b->core; | 
|  | 28 	int i, x, y, u = 0; | 
|  | 29 	kstring_t *str; | 
|  | 30 	int32_t old_nm_i = -1, nm = 0; | 
|  | 31 | 
|  | 32 	str = (kstring_t*)calloc(1, sizeof(kstring_t)); | 
|  | 33 	for (i = y = 0, x = c->pos; i < c->n_cigar; ++i) { | 
|  | 34 		int j, l = cigar[i]>>4, op = cigar[i]&0xf; | 
|  | 35 		if (op == BAM_CMATCH || op == BAM_CEQUAL || op == BAM_CDIFF) { | 
|  | 36 			for (j = 0; j < l; ++j) { | 
|  | 37 				int z = y + j; | 
|  | 38 				int c1 = bam1_seqi(seq, z), c2 = bam_nt16_table[(int)ref[x+j]]; | 
|  | 39 				if (ref[x+j] == 0) break; // out of boundary | 
|  | 40 				if ((c1 == c2 && c1 != 15 && c2 != 15) || c1 == 0) { // a match | 
|  | 41 					if (flag&USE_EQUAL) seq[z/2] &= (z&1)? 0xf0 : 0x0f; | 
|  | 42 					++u; | 
|  | 43 				} else { | 
|  | 44 					kputw(u, str); kputc(ref[x+j], str); | 
|  | 45 					u = 0; ++nm; | 
|  | 46 				} | 
|  | 47 			} | 
|  | 48 			if (j < l) break; | 
|  | 49 			x += l; y += l; | 
|  | 50 		} else if (op == BAM_CDEL) { | 
|  | 51 			kputw(u, str); kputc('^', str); | 
|  | 52 			for (j = 0; j < l; ++j) { | 
|  | 53 				if (ref[x+j] == 0) break; | 
|  | 54 				kputc(ref[x+j], str); | 
|  | 55 			} | 
|  | 56 			u = 0; | 
|  | 57 			if (j < l) break; | 
|  | 58 			x += l; nm += l; | 
|  | 59 		} else if (op == BAM_CINS || op == BAM_CSOFT_CLIP) { | 
|  | 60 			y += l; | 
|  | 61 			if (op == BAM_CINS) nm += l; | 
|  | 62 		} else if (op == BAM_CREF_SKIP) { | 
|  | 63 			x += l; | 
|  | 64 		} | 
|  | 65 	} | 
|  | 66 	kputw(u, str); | 
|  | 67 	// apply max_nm | 
|  | 68 	if (max_nm > 0 && nm >= max_nm) { | 
|  | 69 		for (i = y = 0, x = c->pos; i < c->n_cigar; ++i) { | 
|  | 70 			int j, l = cigar[i]>>4, op = cigar[i]&0xf; | 
|  | 71 			if (op == BAM_CMATCH || op == BAM_CEQUAL || op == BAM_CDIFF) { | 
|  | 72 				for (j = 0; j < l; ++j) { | 
|  | 73 					int z = y + j; | 
|  | 74 					int c1 = bam1_seqi(seq, z), c2 = bam_nt16_table[(int)ref[x+j]]; | 
|  | 75 					if (ref[x+j] == 0) break; // out of boundary | 
|  | 76 					if ((c1 == c2 && c1 != 15 && c2 != 15) || c1 == 0) { // a match | 
|  | 77 						seq[z/2] |= (z&1)? 0x0f : 0xf0; | 
|  | 78 						bam1_qual(b)[z] = 0; | 
|  | 79 					} | 
|  | 80 				} | 
|  | 81 				if (j < l) break; | 
|  | 82 				x += l; y += l; | 
|  | 83 			} else if (op == BAM_CDEL || op == BAM_CREF_SKIP) x += l; | 
|  | 84 			else if (op == BAM_CINS || op == BAM_CSOFT_CLIP) y += l; | 
|  | 85 		} | 
|  | 86 	} | 
|  | 87 	// update NM | 
|  | 88 	if (flag & UPDATE_NM) { | 
|  | 89 		uint8_t *old_nm = bam_aux_get(b, "NM"); | 
|  | 90 		if (c->flag & BAM_FUNMAP) return; | 
|  | 91 		if (old_nm) old_nm_i = bam_aux2i(old_nm); | 
|  | 92 		if (!old_nm) bam_aux_append(b, "NM", 'i', 4, (uint8_t*)&nm); | 
|  | 93 		else if (nm != old_nm_i) { | 
|  | 94 			fprintf(stderr, "[bam_fillmd1] different NM for read '%s': %d -> %d\n", bam1_qname(b), old_nm_i, nm); | 
|  | 95 			bam_aux_del(b, old_nm); | 
|  | 96 			bam_aux_append(b, "NM", 'i', 4, (uint8_t*)&nm); | 
|  | 97 		} | 
|  | 98 	} | 
|  | 99 	// update MD | 
|  | 100 	if (flag & UPDATE_MD) { | 
|  | 101 		uint8_t *old_md = bam_aux_get(b, "MD"); | 
|  | 102 		if (c->flag & BAM_FUNMAP) return; | 
|  | 103 		if (!old_md) bam_aux_append(b, "MD", 'Z', str->l + 1, (uint8_t*)str->s); | 
|  | 104 		else { | 
|  | 105 			int is_diff = 0; | 
|  | 106 			if (strlen((char*)old_md+1) == str->l) { | 
|  | 107 				for (i = 0; i < str->l; ++i) | 
|  | 108 					if (toupper(old_md[i+1]) != toupper(str->s[i])) | 
|  | 109 						break; | 
|  | 110 				if (i < str->l) is_diff = 1; | 
|  | 111 			} else is_diff = 1; | 
|  | 112 			if (is_diff) { | 
|  | 113 				fprintf(stderr, "[bam_fillmd1] different MD for read '%s': '%s' -> '%s'\n", bam1_qname(b), old_md+1, str->s); | 
|  | 114 				bam_aux_del(b, old_md); | 
|  | 115 				bam_aux_append(b, "MD", 'Z', str->l + 1, (uint8_t*)str->s); | 
|  | 116 			} | 
|  | 117 		} | 
|  | 118 	} | 
|  | 119 	// drop all tags but RG | 
|  | 120 	if (flag&DROP_TAG) { | 
|  | 121 		uint8_t *q = bam_aux_get(b, "RG"); | 
|  | 122 		bam_aux_drop_other(b, q); | 
|  | 123 	} | 
|  | 124 	// reduce the resolution of base quality | 
|  | 125 	if (flag&BIN_QUAL) { | 
|  | 126 		uint8_t *qual = bam1_qual(b); | 
|  | 127 		for (i = 0; i < b->core.l_qseq; ++i) | 
|  | 128 			if (qual[i] >= 3) qual[i] = qual[i]/10*10 + 7; | 
|  | 129 	} | 
|  | 130 	free(str->s); free(str); | 
|  | 131 } | 
|  | 132 | 
|  | 133 void bam_fillmd1(bam1_t *b, char *ref, int flag) | 
|  | 134 { | 
|  | 135 	bam_fillmd1_core(b, ref, flag, 0); | 
|  | 136 } | 
|  | 137 | 
|  | 138 int bam_cap_mapQ(bam1_t *b, char *ref, int thres) | 
|  | 139 { | 
|  | 140 	uint8_t *seq = bam1_seq(b), *qual = bam1_qual(b); | 
|  | 141 	uint32_t *cigar = bam1_cigar(b); | 
|  | 142 	bam1_core_t *c = &b->core; | 
|  | 143 	int i, x, y, mm, q, len, clip_l, clip_q; | 
|  | 144 	double t; | 
|  | 145 	if (thres < 0) thres = 40; // set the default | 
|  | 146 	mm = q = len = clip_l = clip_q = 0; | 
|  | 147 	for (i = y = 0, x = c->pos; i < c->n_cigar; ++i) { | 
|  | 148 		int j, l = cigar[i]>>4, op = cigar[i]&0xf; | 
|  | 149 		if (op == BAM_CMATCH || op == BAM_CEQUAL || op == BAM_CDIFF) { | 
|  | 150 			for (j = 0; j < l; ++j) { | 
|  | 151 				int z = y + j; | 
|  | 152 				int c1 = bam1_seqi(seq, z), c2 = bam_nt16_table[(int)ref[x+j]]; | 
|  | 153 				if (ref[x+j] == 0) break; // out of boundary | 
|  | 154 				if (c2 != 15 && c1 != 15 && qual[z] >= 13) { // not ambiguous | 
|  | 155 					++len; | 
|  | 156 					if (c1 && c1 != c2 && qual[z] >= 13) { // mismatch | 
|  | 157 						++mm; | 
|  | 158 						q += qual[z] > 33? 33 : qual[z]; | 
|  | 159 					} | 
|  | 160 				} | 
|  | 161 			} | 
|  | 162 			if (j < l) break; | 
|  | 163 			x += l; y += l; len += l; | 
|  | 164 		} else if (op == BAM_CDEL) { | 
|  | 165 			for (j = 0; j < l; ++j) | 
|  | 166 				if (ref[x+j] == 0) break; | 
|  | 167 			if (j < l) break; | 
|  | 168 			x += l; | 
|  | 169 		} else if (op == BAM_CSOFT_CLIP) { | 
|  | 170 			for (j = 0; j < l; ++j) clip_q += qual[y+j]; | 
|  | 171 			clip_l += l; | 
|  | 172 			y += l; | 
|  | 173 		} else if (op == BAM_CHARD_CLIP) { | 
|  | 174 			clip_q += 13 * l; | 
|  | 175 			clip_l += l; | 
|  | 176 		} else if (op == BAM_CINS) y += l; | 
|  | 177 		else if (op == BAM_CREF_SKIP) x += l; | 
|  | 178 	} | 
|  | 179 	for (i = 0, t = 1; i < mm; ++i) | 
|  | 180 		t *= (double)len / (i+1); | 
|  | 181 	t = q - 4.343 * log(t) + clip_q / 5.; | 
|  | 182 	if (t > thres) return -1; | 
|  | 183 	if (t < 0) t = 0; | 
|  | 184 	t = sqrt((thres - t) / thres) * thres; | 
|  | 185 //	fprintf(stderr, "%s %lf %d\n", bam1_qname(b), t, q); | 
|  | 186 	return (int)(t + .499); | 
|  | 187 } | 
|  | 188 | 
|  | 189 int bam_prob_realn_core(bam1_t *b, const char *ref, int flag) | 
|  | 190 { | 
|  | 191 	int k, i, bw, x, y, yb, ye, xb, xe, apply_baq = flag&1, extend_baq = flag>>1&1, redo_baq = flag&4; | 
|  | 192 	uint32_t *cigar = bam1_cigar(b); | 
|  | 193 	bam1_core_t *c = &b->core; | 
|  | 194 	kpa_par_t conf = kpa_par_def; | 
|  | 195 	uint8_t *bq = 0, *zq = 0, *qual = bam1_qual(b); | 
|  | 196 	if ((c->flag & BAM_FUNMAP) || b->core.l_qseq == 0) return -1; // do nothing | 
|  | 197 	// test if BQ or ZQ is present | 
|  | 198 	if ((bq = bam_aux_get(b, "BQ")) != 0) ++bq; | 
|  | 199 	if ((zq = bam_aux_get(b, "ZQ")) != 0 && *zq == 'Z') ++zq; | 
|  | 200 	if (bq && redo_baq) | 
|  | 201 	{ | 
|  | 202 	    bam_aux_del(b, bq-1); | 
|  | 203 	    bq = 0; | 
|  | 204 	} | 
|  | 205 	if (bq && zq) { // remove the ZQ tag | 
|  | 206 		bam_aux_del(b, zq-1); | 
|  | 207 		zq = 0; | 
|  | 208 	} | 
|  | 209 	if (bq || zq) { | 
|  | 210 		if ((apply_baq && zq) || (!apply_baq && bq)) return -3; // in both cases, do nothing | 
|  | 211 		if (bq && apply_baq) { // then convert BQ to ZQ | 
|  | 212 			for (i = 0; i < c->l_qseq; ++i) | 
|  | 213 				qual[i] = qual[i] + 64 < bq[i]? 0 : qual[i] - ((int)bq[i] - 64); | 
|  | 214 			*(bq - 3) = 'Z'; | 
|  | 215 		} else if (zq && !apply_baq) { // then convert ZQ to BQ | 
|  | 216 			for (i = 0; i < c->l_qseq; ++i) | 
|  | 217 				qual[i] += (int)zq[i] - 64; | 
|  | 218 			*(zq - 3) = 'B'; | 
|  | 219 		} | 
|  | 220 		return 0; | 
|  | 221 	} | 
|  | 222 	// find the start and end of the alignment | 
|  | 223 	x = c->pos, y = 0, yb = ye = xb = xe = -1; | 
|  | 224 	for (k = 0; k < c->n_cigar; ++k) { | 
|  | 225 		int op, l; | 
|  | 226 		op = cigar[k]&0xf; l = cigar[k]>>4; | 
|  | 227 		if (op == BAM_CMATCH || op == BAM_CEQUAL || op == BAM_CDIFF) { | 
|  | 228 			if (yb < 0) yb = y; | 
|  | 229 			if (xb < 0) xb = x; | 
|  | 230 			ye = y + l; xe = x + l; | 
|  | 231 			x += l; y += l; | 
|  | 232 		} else if (op == BAM_CSOFT_CLIP || op == BAM_CINS) y += l; | 
|  | 233 		else if (op == BAM_CDEL) x += l; | 
|  | 234 		else if (op == BAM_CREF_SKIP) return -1; // do nothing if there is a reference skip | 
|  | 235 	} | 
|  | 236 	// set bandwidth and the start and the end | 
|  | 237 	bw = 7; | 
|  | 238 	if (abs((xe - xb) - (ye - yb)) > bw) | 
|  | 239 		bw = abs((xe - xb) - (ye - yb)) + 3; | 
|  | 240 	conf.bw = bw; | 
|  | 241 	xb -= yb + bw/2; if (xb < 0) xb = 0; | 
|  | 242 	xe += c->l_qseq - ye + bw/2; | 
|  | 243 	if (xe - xb - c->l_qseq > bw) | 
|  | 244 		xb += (xe - xb - c->l_qseq - bw) / 2, xe -= (xe - xb - c->l_qseq - bw) / 2; | 
|  | 245 	{ // glocal | 
|  | 246 		uint8_t *s, *r, *q, *seq = bam1_seq(b), *bq; | 
|  | 247 		int *state; | 
|  | 248 		bq = calloc(c->l_qseq + 1, 1); | 
|  | 249 		memcpy(bq, qual, c->l_qseq); | 
|  | 250 		s = calloc(c->l_qseq, 1); | 
|  | 251 		for (i = 0; i < c->l_qseq; ++i) s[i] = bam_nt16_nt4_table[bam1_seqi(seq, i)]; | 
|  | 252 		r = calloc(xe - xb, 1); | 
|  | 253 		for (i = xb; i < xe; ++i) { | 
|  | 254 			if (ref[i] == 0) { xe = i; break; } | 
|  | 255 			r[i-xb] = bam_nt16_nt4_table[bam_nt16_table[(int)ref[i]]]; | 
|  | 256 		} | 
|  | 257 		state = calloc(c->l_qseq, sizeof(int)); | 
|  | 258 		q = calloc(c->l_qseq, 1); | 
|  | 259 		kpa_glocal(r, xe-xb, s, c->l_qseq, qual, &conf, state, q); | 
|  | 260 		if (!extend_baq) { // in this block, bq[] is capped by base quality qual[] | 
|  | 261 			for (k = 0, x = c->pos, y = 0; k < c->n_cigar; ++k) { | 
|  | 262 				int op = cigar[k]&0xf, l = cigar[k]>>4; | 
|  | 263 				if (op == BAM_CMATCH || op == BAM_CEQUAL || op == BAM_CDIFF) { | 
|  | 264 					for (i = y; i < y + l; ++i) { | 
|  | 265 						if ((state[i]&3) != 0 || state[i]>>2 != x - xb + (i - y)) bq[i] = 0; | 
|  | 266 						else bq[i] = bq[i] < q[i]? bq[i] : q[i]; | 
|  | 267 					} | 
|  | 268 					x += l; y += l; | 
|  | 269 				} else if (op == BAM_CSOFT_CLIP || op == BAM_CINS) y += l; | 
|  | 270 				else if (op == BAM_CDEL) x += l; | 
|  | 271 			} | 
|  | 272 			for (i = 0; i < c->l_qseq; ++i) bq[i] = qual[i] - bq[i] + 64; // finalize BQ | 
|  | 273 		} else { // in this block, bq[] is BAQ that can be larger than qual[] (different from the above!) | 
|  | 274 			uint8_t *left, *rght; | 
|  | 275 			left = calloc(c->l_qseq, 1); rght = calloc(c->l_qseq, 1); | 
|  | 276 			for (k = 0, x = c->pos, y = 0; k < c->n_cigar; ++k) { | 
|  | 277 				int op = cigar[k]&0xf, l = cigar[k]>>4; | 
|  | 278 				if (op == BAM_CMATCH || op == BAM_CEQUAL || op == BAM_CDIFF) { | 
|  | 279 					for (i = y; i < y + l; ++i) | 
|  | 280 						bq[i] = ((state[i]&3) != 0 || state[i]>>2 != x - xb + (i - y))? 0 : q[i]; | 
|  | 281 					for (left[y] = bq[y], i = y + 1; i < y + l; ++i) | 
|  | 282 						left[i] = bq[i] > left[i-1]? bq[i] : left[i-1]; | 
|  | 283 					for (rght[y+l-1] = bq[y+l-1], i = y + l - 2; i >= y; --i) | 
|  | 284 						rght[i] = bq[i] > rght[i+1]? bq[i] : rght[i+1]; | 
|  | 285 					for (i = y; i < y + l; ++i) | 
|  | 286 						bq[i] = left[i] < rght[i]? left[i] : rght[i]; | 
|  | 287 					x += l; y += l; | 
|  | 288 				} else if (op == BAM_CSOFT_CLIP || op == BAM_CINS) y += l; | 
|  | 289 				else if (op == BAM_CDEL) x += l; | 
|  | 290 			} | 
|  | 291 			for (i = 0; i < c->l_qseq; ++i) bq[i] = 64 + (qual[i] <= bq[i]? 0 : qual[i] - bq[i]); // finalize BQ | 
|  | 292 			free(left); free(rght); | 
|  | 293 		} | 
|  | 294 		if (apply_baq) { | 
|  | 295 			for (i = 0; i < c->l_qseq; ++i) qual[i] -= bq[i] - 64; // modify qual | 
|  | 296 			bam_aux_append(b, "ZQ", 'Z', c->l_qseq + 1, bq); | 
|  | 297 		} else bam_aux_append(b, "BQ", 'Z', c->l_qseq + 1, bq); | 
|  | 298 		free(bq); free(s); free(r); free(q); free(state); | 
|  | 299 	} | 
|  | 300 	return 0; | 
|  | 301 } | 
|  | 302 | 
|  | 303 int bam_prob_realn(bam1_t *b, const char *ref) | 
|  | 304 { | 
|  | 305 	return bam_prob_realn_core(b, ref, 1); | 
|  | 306 } | 
|  | 307 | 
|  | 308 int bam_fillmd(int argc, char *argv[]) | 
|  | 309 { | 
|  | 310 	int c, flt_flag, tid = -2, ret, len, is_bam_out, is_sam_in, is_uncompressed, max_nm, is_realn, capQ, baq_flag; | 
|  | 311 	samfile_t *fp, *fpout = 0; | 
|  | 312 	faidx_t *fai; | 
|  | 313 	char *ref = 0, mode_w[8], mode_r[8]; | 
|  | 314 	bam1_t *b; | 
|  | 315 | 
|  | 316 	flt_flag = UPDATE_NM | UPDATE_MD; | 
|  | 317 	is_bam_out = is_sam_in = is_uncompressed = is_realn = max_nm = capQ = baq_flag = 0; | 
|  | 318 	mode_w[0] = mode_r[0] = 0; | 
|  | 319 	strcpy(mode_r, "r"); strcpy(mode_w, "w"); | 
|  | 320 	while ((c = getopt(argc, argv, "EqreuNhbSC:n:Ad")) >= 0) { | 
|  | 321 		switch (c) { | 
|  | 322 		case 'r': is_realn = 1; break; | 
|  | 323 		case 'e': flt_flag |= USE_EQUAL; break; | 
|  | 324 		case 'd': flt_flag |= DROP_TAG; break; | 
|  | 325 		case 'q': flt_flag |= BIN_QUAL; break; | 
|  | 326 		case 'h': flt_flag |= HASH_QNM; break; | 
|  | 327 		case 'N': flt_flag &= ~(UPDATE_MD|UPDATE_NM); break; | 
|  | 328 		case 'b': is_bam_out = 1; break; | 
|  | 329 		case 'u': is_uncompressed = is_bam_out = 1; break; | 
|  | 330 		case 'S': is_sam_in = 1; break; | 
|  | 331 		case 'n': max_nm = atoi(optarg); break; | 
|  | 332 		case 'C': capQ = atoi(optarg); break; | 
|  | 333 		case 'A': baq_flag |= 1; break; | 
|  | 334 		case 'E': baq_flag |= 2; break; | 
|  | 335 		default: fprintf(stderr, "[bam_fillmd] unrecognized option '-%c'\n", c); return 1; | 
|  | 336 		} | 
|  | 337 	} | 
|  | 338 	if (!is_sam_in) strcat(mode_r, "b"); | 
|  | 339 	if (is_bam_out) strcat(mode_w, "b"); | 
|  | 340 	else strcat(mode_w, "h"); | 
|  | 341 	if (is_uncompressed) strcat(mode_w, "u"); | 
|  | 342 	if (optind + 1 >= argc) { | 
|  | 343 		fprintf(stderr, "\n"); | 
|  | 344 		fprintf(stderr, "Usage:   samtools fillmd [-eubrS] <aln.bam> <ref.fasta>\n\n"); | 
|  | 345 		fprintf(stderr, "Options: -e       change identical bases to '='\n"); | 
|  | 346 		fprintf(stderr, "         -u       uncompressed BAM output (for piping)\n"); | 
|  | 347 		fprintf(stderr, "         -b       compressed BAM output\n"); | 
|  | 348 		fprintf(stderr, "         -S       the input is SAM with header\n"); | 
|  | 349 		fprintf(stderr, "         -A       modify the quality string\n"); | 
|  | 350 		fprintf(stderr, "         -r       compute the BQ tag (without -A) or cap baseQ by BAQ (with -A)\n"); | 
|  | 351 		fprintf(stderr, "         -E       extended BAQ for better sensitivity but lower specificity\n\n"); | 
|  | 352 		return 1; | 
|  | 353 	} | 
|  | 354 	fp = samopen(argv[optind], mode_r, 0); | 
|  | 355 	if (fp == 0) return 1; | 
|  | 356 	if (is_sam_in && (fp->header == 0 || fp->header->n_targets == 0)) { | 
|  | 357 		fprintf(stderr, "[bam_fillmd] input SAM does not have header. Abort!\n"); | 
|  | 358 		return 1; | 
|  | 359 	} | 
|  | 360 	fpout = samopen("-", mode_w, fp->header); | 
|  | 361 	fai = fai_load(argv[optind+1]); | 
|  | 362 | 
|  | 363 	b = bam_init1(); | 
|  | 364 	while ((ret = samread(fp, b)) >= 0) { | 
|  | 365 		if (b->core.tid >= 0) { | 
|  | 366 			if (tid != b->core.tid) { | 
|  | 367 				free(ref); | 
|  | 368 				ref = fai_fetch(fai, fp->header->target_name[b->core.tid], &len); | 
|  | 369 				tid = b->core.tid; | 
|  | 370 				if (ref == 0) | 
|  | 371 					fprintf(stderr, "[bam_fillmd] fail to find sequence '%s' in the reference.\n", | 
|  | 372 							fp->header->target_name[tid]); | 
|  | 373 			} | 
|  | 374 			if (is_realn) bam_prob_realn_core(b, ref, baq_flag); | 
|  | 375 			if (capQ > 10) { | 
|  | 376 				int q = bam_cap_mapQ(b, ref, capQ); | 
|  | 377 				if (b->core.qual > q) b->core.qual = q; | 
|  | 378 			} | 
|  | 379 			if (ref) bam_fillmd1_core(b, ref, flt_flag, max_nm); | 
|  | 380 		} | 
|  | 381 		samwrite(fpout, b); | 
|  | 382 	} | 
|  | 383 	bam_destroy1(b); | 
|  | 384 | 
|  | 385 	free(ref); | 
|  | 386 	fai_destroy(fai); | 
|  | 387 	samclose(fp); samclose(fpout); | 
|  | 388 	return 0; | 
|  | 389 } |