]> git.donarmstrong.com Git - samtools.git/blobdiff - bam_plcmd.c
minor changes
[samtools.git] / bam_plcmd.c
index e2601ecc3c26b41051d0148a9cf9696036d297fb..195a59602ab4e778084b6eefc105ee1d608643bb 100644 (file)
@@ -5,6 +5,7 @@
 #include "sam.h"
 #include "faidx.h"
 #include "bam_maqcns.h"
+#include "bam_mcns.h"
 #include "khash.h"
 #include "glf.h"
 #include "kstring.h"
@@ -18,6 +19,10 @@ KHASH_MAP_INIT_INT64(64, indel_list_t)
 #define BAM_PLF_GLF        0x08
 #define BAM_PLF_VAR_ONLY   0x10
 #define BAM_PLF_2ND        0x20
+#define BAM_PLF_RANBASE    0x40
+#define BAM_PLF_1STBASE    0x80
+#define BAM_PLF_ALLBASE    0x100
+#define BAM_PLF_READPOS    0x200
 
 typedef struct {
        bam_header_t *h;
@@ -28,6 +33,7 @@ typedef struct {
        uint32_t format;
        int tid, len, last_pos;
        int mask;
+    int max_depth;  // for indel calling, ignore reads with the depth too high. 0 for unlimited
        char *ref;
        glfFile fp_glf; // for glf output only
 } pu_data_t;
@@ -121,9 +127,12 @@ static int glt3_func(uint32_t tid, uint32_t pos, int n, const bam_pileup1_t *pu,
        g3->offset = pos - d->last_pos;
        d->last_pos = pos;
        glf3_write1(d->fp_glf, g3);
-       if (proposed_indels)
-               r = bam_maqindel(n, pos, d->ido, pu, d->ref, proposed_indels[0], proposed_indels+1);
-       else r = bam_maqindel(n, pos, d->ido, pu, d->ref, 0, 0);
+    if (pos < d->len) {
+        int m = (!d->max_depth || d->max_depth>n) ? n : d->max_depth;
+               if (proposed_indels)
+                       r = bam_maqindel(m, pos, d->ido, pu, d->ref, proposed_indels[0], proposed_indels+1);
+               else r = bam_maqindel(m, pos, d->ido, pu, d->ref, 0, 0);
+       }
        if (r) { // then write indel line
                int het = 3 * n, min;
                min = het;
@@ -150,11 +159,37 @@ static int glt3_func(uint32_t tid, uint32_t pos, int n, const bam_pileup1_t *pu,
        return 0;
 }
 
+static void pileup_seq(const bam_pileup1_t *p, int pos, int ref_len, const char *ref)
+{
+       if (p->is_head) printf("^%c", p->b->core.qual > 93? 126 : p->b->core.qual + 33);
+       if (!p->is_del) {
+               int j, rb, c = bam_nt16_rev_table[bam1_seqi(bam1_seq(p->b), p->qpos)];
+               rb = (ref && pos < ref_len)? ref[pos] : 'N';
+               if (c == '=' || toupper(c) == toupper(rb)) c = bam1_strand(p->b)? ',' : '.';
+               else c = bam1_strand(p->b)? tolower(c) : toupper(c);
+               putchar(c);
+               if (p->indel > 0) {
+                       printf("+%d", p->indel);
+                       for (j = 1; j <= p->indel; ++j) {
+                               c = bam_nt16_rev_table[bam1_seqi(bam1_seq(p->b), p->qpos + j)];
+                               putchar(bam1_strand(p->b)? tolower(c) : toupper(c));
+                       }
+               } else if (p->indel < 0) {
+                       printf("%d", p->indel);
+                       for (j = 1; j <= -p->indel; ++j) {
+                               c = (ref && (int)pos+j < ref_len)? ref[pos+j] : 'N';
+                               putchar(bam1_strand(p->b)? tolower(c) : toupper(c));
+                       }
+               }
+       } else putchar('*');
+       if (p->is_tail) putchar('$');
+}
+
 static int pileup_func(uint32_t tid, uint32_t pos, int n, const bam_pileup1_t *pu, void *data)
 {
        pu_data_t *d = (pu_data_t*)data;
        bam_maqindel_ret_t *r = 0;
-       int i, j, rb, rms_mapq = -1, *proposed_indels = 0;
+       int i, rb, rms_mapq = -1, *proposed_indels = 0;
        uint64_t rms_aux;
        uint32_t cns = 0;
 
@@ -169,7 +204,7 @@ static int pileup_func(uint32_t tid, uint32_t pos, int n, const bam_pileup1_t *p
        // update d->ref if necessary
        if (d->fai && (int)tid != d->tid) {
                free(d->ref);
-               d->ref = fai_fetch(d->fai, d->h->target_name[tid], &d->len);
+               d->ref = faidx_fetch_seq(d->fai, d->h->target_name[tid], 0, 0x7fffffff, &d->len);
                d->tid = tid;
        }
        rb = (d->ref && (int)pos < d->len)? d->ref[pos] : 'N';
@@ -180,12 +215,31 @@ static int pileup_func(uint32_t tid, uint32_t pos, int n, const bam_pileup1_t *p
                if (i == n) return 0;
        }
        // call the consensus and indel
-       if (d->format & BAM_PLF_CNS) // call consensus
-               cns = bam_maqcns_call(n, pu, d->c);
-       if ((d->format & (BAM_PLF_CNS|BAM_PLF_INDEL_ONLY)) && d->ref && pos < d->len) { // call indels
-               if (proposed_indels) // the first element gives the size of the array
-                       r = bam_maqindel(n, pos, d->ido, pu, d->ref, proposed_indels[0], proposed_indels+1);
-               else r = bam_maqindel(n, pos, d->ido, pu, d->ref, 0, 0);
+       if (d->format & BAM_PLF_CNS) { // call consensus
+               if (d->format & (BAM_PLF_RANBASE|BAM_PLF_1STBASE)) { // use a random base or the 1st base as the consensus call
+                       const bam_pileup1_t *p = (d->format & BAM_PLF_1STBASE)? pu : pu + (int)(drand48() * n);
+                       int q = bam1_qual(p->b)[p->qpos];
+                       int mapQ = p->b->core.qual < d->c->cap_mapQ? p->b->core.qual : d->c->cap_mapQ;
+                       uint32_t b = bam1_seqi(bam1_seq(p->b), p->qpos);
+                       cns = b<<28 | 0xf<<24 | mapQ<<16 | q<<8;
+               } else if (d->format & BAM_PLF_ALLBASE) { // collapse all bases
+                       uint64_t rmsQ = 0;
+                       uint32_t b = 0;
+                       for (i = 0; i < n; ++i) {
+                               const bam_pileup1_t *p = pu + i;
+                               int q = p->b->core.qual < d->c->cap_mapQ? p->b->core.qual : d->c->cap_mapQ;
+                               b |= bam1_seqi(bam1_seq(p->b), p->qpos);
+                               rmsQ += q * q;
+                       }
+                       rmsQ = (uint64_t)(sqrt((double)rmsQ / n) + .499);
+                       cns = b<<28 | 0xf<<24 | rmsQ<<16 | 60<<8;
+               } else cns = bam_maqcns_call(n, pu, d->c);
+       }
+    if ((d->format & (BAM_PLF_CNS|BAM_PLF_INDEL_ONLY)) && d->ref && pos < d->len) { // call indels
+        int m = (!d->max_depth || d->max_depth>n) ? n : d->max_depth;
+        if (proposed_indels) // the first element gives the size of the array
+            r = bam_maqindel(m, pos, d->ido, pu, d->ref, proposed_indels[0], proposed_indels+1);
+        else r = bam_maqindel(m, pos, d->ido, pu, d->ref, 0, 0);
        }
        // when only variant sites are asked for, test if the site is a variant
        if ((d->format & BAM_PLF_CNS) && (d->format & BAM_PLF_VAR_ONLY)) {
@@ -216,27 +270,7 @@ static int pileup_func(uint32_t tid, uint32_t pos, int n, const bam_pileup1_t *p
                const bam_pileup1_t *p = pu + i;
                int tmp = p->b->core.qual < d->c->cap_mapQ? p->b->core.qual : d->c->cap_mapQ;
                rms_aux += tmp * tmp;
-               if (p->is_head) printf("^%c", p->b->core.qual > 93? 126 : p->b->core.qual + 33);
-               if (!p->is_del) {
-                       int c = bam_nt16_rev_table[bam1_seqi(bam1_seq(p->b), p->qpos)];
-                       if (c == '=' || toupper(c) == toupper(rb)) c = bam1_strand(p->b)? ',' : '.';
-                       else c = bam1_strand(p->b)? tolower(c) : toupper(c);
-                       putchar(c);
-                       if (p->indel > 0) {
-                               printf("+%d", p->indel);
-                               for (j = 1; j <= p->indel; ++j) {
-                                       c = bam_nt16_rev_table[bam1_seqi(bam1_seq(p->b), p->qpos + j)];
-                                       putchar(bam1_strand(p->b)? tolower(c) : toupper(c));
-                               }
-                       } else if (p->indel < 0) {
-                               printf("%d", p->indel);
-                               for (j = 1; j <= -p->indel; ++j) {
-                                       c = (d->ref && (int)pos+j < d->len)? d->ref[pos+j] : 'N';
-                                       putchar(bam1_strand(p->b)? tolower(c) : toupper(c));
-                               }
-                       }
-               } else putchar('*');
-               if (p->is_tail) putchar('$');
+               pileup_seq(p, pos, d->len, d->ref);
        }
        // finalize rms_mapq
        rms_aux = (uint64_t)(sqrt((double)rms_aux / n) + .499);
@@ -273,6 +307,15 @@ static int pileup_func(uint32_t tid, uint32_t pos, int n, const bam_pileup1_t *p
                        putchar(c);
                }
        }
+       // print read position
+       if (d->format & BAM_PLF_READPOS) {
+               putchar('\t');
+               for (i = 0; i < n; ++i) {
+                       int x = pu[i].qpos;
+                       int l = pu[i].b->core.l_qseq;
+                       printf("%d,", x < l/2? x+1 : -((l-1)-x+1));
+               }
+       }
        putchar('\n');
        // print the indel line if r has been calculated. This only happens if:
        // a) -c or -i are flagged, AND b) the reference sequence is available
@@ -296,29 +339,40 @@ int bam_pileup(int argc, char *argv[])
        int c, is_SAM = 0;
        char *fn_list = 0, *fn_fa = 0, *fn_pos = 0;
        pu_data_t *d = (pu_data_t*)calloc(1, sizeof(pu_data_t));
+    d->max_depth = 0;
        d->tid = -1; d->mask = BAM_DEF_MASK;
        d->c = bam_maqcns_init();
+       d->c->is_soap = 1; // change the default model
        d->ido = bam_maqindel_opt_init();
-       while ((c = getopt(argc, argv, "st:f:cT:N:r:l:im:gI:G:vM:S2a")) >= 0) {
+       while ((c = getopt(argc, argv, "st:f:cT:N:r:l:d:im:gI:G:vM:S2aR:PA")) >= 0) {
                switch (c) {
                case 'a': d->c->is_soap = 1; break;
+               case 'A': d->c->is_soap = 0; break;
                case 's': d->format |= BAM_PLF_SIMPLE; break;
                case 't': fn_list = strdup(optarg); break;
                case 'l': fn_pos = strdup(optarg); break;
                case 'f': fn_fa = strdup(optarg); break;
                case 'T': d->c->theta = atof(optarg); break;
                case 'N': d->c->n_hap = atoi(optarg); break;
-               case 'r': d->c->het_rate = atof(optarg); break;
+               case 'r': d->c->het_rate = atof(optarg); d->ido->r_snp = d->c->het_rate; break;
                case 'M': d->c->cap_mapQ = atoi(optarg); break;
+               case 'd': d->max_depth = atoi(optarg); break;
                case 'c': d->format |= BAM_PLF_CNS; break;
                case 'i': d->format |= BAM_PLF_INDEL_ONLY; break;
                case 'v': d->format |= BAM_PLF_VAR_ONLY; break;
                case 'm': d->mask = strtol(optarg, 0, 0); break;
                case 'g': d->format |= BAM_PLF_GLF; break;
                case '2': d->format |= BAM_PLF_2ND; break;
+               case 'P': d->format |= BAM_PLF_READPOS; break;
                case 'I': d->ido->q_indel = atoi(optarg); break;
                case 'G': d->ido->r_indel = atof(optarg); break;
                case 'S': is_SAM = 1; break;
+               case 'R':
+                       if (strcmp(optarg, "random") == 0) d->format |= BAM_PLF_RANBASE;
+                       else if (strcmp(optarg, "first") == 0) d->format |= BAM_PLF_1STBASE;
+                       else if (strcmp(optarg, "all") == 0) d->format |= BAM_PLF_ALLBASE;
+                       else fprintf(stderr, "[bam_pileup] unrecognized -R\n");
+                       break;
                default: fprintf(stderr, "Unrecognizd option '-%c'.\n", c); return 1;
                }
        }
@@ -328,15 +382,16 @@ int bam_pileup(int argc, char *argv[])
                fprintf(stderr, "Usage:  samtools pileup [options] <in.bam>|<in.sam>\n\n");
                fprintf(stderr, "Option: -s        simple (yet incomplete) pileup format\n");
                fprintf(stderr, "        -S        the input is in SAM\n");
-               fprintf(stderr, "        -a        use the SOAPsnp model for SNP calling\n");
+               fprintf(stderr, "        -A        use the MAQ model for SNP calling\n");
                fprintf(stderr, "        -2        output the 2nd best call and quality\n");
                fprintf(stderr, "        -i        only show lines/consensus with indels\n");
                fprintf(stderr, "        -m INT    filtering reads with bits in INT [%d]\n", d->mask);
                fprintf(stderr, "        -M INT    cap mapping quality at INT [%d]\n", d->c->cap_mapQ);
+        fprintf(stderr, "        -d INT    limit maximum depth for indels [unlimited]\n");
                fprintf(stderr, "        -t FILE   list of reference sequences (force -S)\n");
                fprintf(stderr, "        -l FILE   list of sites at which pileup is output\n");
                fprintf(stderr, "        -f FILE   reference sequence in the FASTA format\n\n");
-               fprintf(stderr, "        -c        output the maq consensus sequence\n");
+               fprintf(stderr, "        -c        output the SOAPsnp consensus sequence\n");
                fprintf(stderr, "        -v        print variants only (for -c)\n");
                fprintf(stderr, "        -g        output in the GLFv3 format (suppressing -c/-i/-s)\n");
                fprintf(stderr, "        -T FLOAT  theta in maq consensus calling model (for -c/-g) [%f]\n", d->c->theta);
@@ -348,6 +403,7 @@ int bam_pileup(int argc, char *argv[])
                free(fn_list); free(fn_fa); free(d);
                return 1;
        }
+       if (d->format & (BAM_PLF_RANBASE|BAM_PLF_1STBASE|BAM_PLF_ALLBASE)) d->format |= BAM_PLF_CNS;
        if (fn_fa) d->fai = fai_load(fn_fa);
        if (d->format & (BAM_PLF_CNS|BAM_PLF_GLF)) bam_maqcns_prepare(d->c); // consensus calling
        if (d->format & BAM_PLF_GLF) { // for glf output
@@ -388,3 +444,265 @@ int bam_pileup(int argc, char *argv[])
        free(d->ido); free(d->ref); free(d);
        return 0;
 }
+
+/***********
+ * mpileup *
+ ***********/
+
+#define MPLP_VCF   0x1
+#define MPLP_VAR   0x2
+#define MPLP_AFS   0x4
+#define MPLP_AFALL 0x8
+
+#define MPLP_AFS_BLOCK 0x10000
+
+typedef struct {
+       int max_mq, min_mq, prior_type, flag;
+       double theta;
+       char *reg, *fn_pos;
+       faidx_t *fai;
+       kh_64_t *hash;
+} mplp_conf_t;
+
+typedef struct {
+       bamFile fp;
+       bam_iter_t iter;
+       int min_mq;
+} mplp_aux_t;
+
+static int mplp_func(void *data, bam1_t *b)
+{
+       mplp_aux_t *ma = (mplp_aux_t*)data;
+       int ret;
+       do {
+               ret = ma->iter? bam_iter_read(ma->fp, ma->iter, b) : bam_read1(ma->fp, b);
+       } while (b->core.qual < ma->min_mq && ret >= 0);
+       return ret;
+}
+
+static int mpileup(mplp_conf_t *conf, int n, char **fn)
+{
+       mplp_aux_t **data;
+       mc_aux_t *ma = 0;
+       int i, tid, pos, *n_plp, beg0 = 0, end0 = 1u<<29, ref_len, ref_tid;
+       const bam_pileup1_t **plp;
+       bam_mplp_t iter;
+       bam_header_t *h = 0;
+       uint64_t N = 0;
+       char *ref;
+       khash_t(64) *hash = 0;
+       // allocate
+       data = calloc(n, sizeof(void*));
+       plp = calloc(n, sizeof(void*));
+       n_plp = calloc(n, sizeof(int*));
+       // read the header and initialize data
+       for (i = 0; i < n; ++i) {
+               bam_header_t *h_tmp;
+               data[i] = calloc(1, sizeof(mplp_aux_t));
+               data[i]->min_mq = conf->min_mq;
+               data[i]->fp = bam_open(fn[i], "r");
+               h_tmp = bam_header_read(data[i]->fp);
+               if (conf->reg) {
+                       int beg, end;
+                       bam_index_t *idx;
+                       idx = bam_index_load(fn[i]);
+                       if (idx == 0) {
+                               fprintf(stderr, "[%s] fail to load index for %d-th input.\n", __func__, i+1);
+                               exit(1);
+                       }
+                       if (bam_parse_region(h_tmp, conf->reg, &tid, &beg, &end) < 0) {
+                               fprintf(stderr, "[%s] malformatted region or wrong seqname for %d-th input.\n", __func__, i+1);
+                               exit(1);
+                       }
+                       if (i == 0) beg0 = beg, end0 = end;
+                       data[i]->iter = bam_iter_query(idx, tid, beg, end);
+                       bam_index_destroy(idx);
+               }
+               if (i == 0) h = h_tmp;
+               else {
+                       // FIXME: to check consistency
+                       bam_header_destroy(h_tmp);
+               }
+       }
+       if (conf->fn_pos) hash = load_pos(conf->fn_pos, h);
+       // write the VCF header
+       if (conf->flag & MPLP_VCF) {
+               kstring_t s;
+               s.l = s.m = 0; s.s = 0;
+               puts("##fileformat=VCFv4.0");
+               puts("##INFO=<ID=DP,Number=1,Type=Integer,Description=\"Total read depth\">");
+               puts("##INFO=<ID=AF,Number=1,Type=Float,Description=\"Non-reference allele frequency \\argmax_f P(D|f)\">");
+               puts("##INFO=<ID=AFE,Number=1,Type=Float,Description=\"Expected non-reference allele frequency\">");
+               puts("##FILTER=<ID=Q13,Description=\"All min{baseQ,mapQ} below 13\">");
+               kputs("#CHROM\tPOS\tID\tREF\tALT\tQUAL\tFILTER\tINFO\tFORMAT", &s);
+               for (i = 0; i < n; ++i) {
+                       const char *p;
+                       kputc('\t', &s);
+                       if ((p = strstr(fn[i], ".bam")) != 0)
+                               kputsn(fn[i], p - fn[i], &s);
+                       else kputs(fn[i], &s);
+               }
+               puts(s.s);
+               free(s.s);
+       }
+       // mpileup
+       if (conf->flag & MPLP_VCF) {
+               ma = mc_init(n);
+               mc_init_prior(ma, conf->prior_type, conf->theta);
+       }
+       ref_tid = -1; ref = 0;
+       iter = bam_mplp_init(n, mplp_func, (void**)data);
+       while (bam_mplp_auto(iter, &tid, &pos, n_plp, plp) > 0) {
+               if (conf->reg && (pos < beg0 || pos >= end0)) continue; // out of the region requested
+               if (hash) {
+                       khint_t k;
+                       k = kh_get(64, hash, (uint64_t)tid<<32 | pos);
+                       if (k == kh_end(hash)) continue;
+               }
+               if (tid != ref_tid) {
+                       free(ref);
+                       if (conf->fai) ref = fai_fetch(conf->fai, h->target_name[tid], &ref_len);
+                       ref_tid = tid;
+               }
+               if (conf->flag & MPLP_VCF) {
+                       mc_rst_t r;
+                       int j, _ref0, depth, rms_q, _ref0b, is_var = 0, qref = 0, level = 2, tot;
+                       uint64_t sqr_sum;
+                       if (conf->flag & MPLP_AFS) level = 3;
+                       _ref0 = _ref0b = (ref && pos < ref_len)? ref[pos] : 'N';
+                       _ref0 = bam_nt16_nt4_table[bam_nt16_table[_ref0]];
+                       tot = mc_cal(_ref0, n_plp, plp, ma, &r, level);
+                       if (tot) { // has good bases
+                               double q;
+                               is_var = (r.p_ref < .5);
+                               q = is_var? r.p_ref : 1. - r.p_ref;
+                               if (q < 1e-308) q = 1e-308;
+                               qref = (int)(-3.434 * log(q) + .499);
+                               if (qref > 99) qref = 99;
+                       }
+                       if ((conf->flag & MPLP_VAR) && !is_var) continue;
+                       ++N; // number of processed lines
+                       printf("%s\t%d\t.\t%c\t", h->target_name[tid], pos + 1, _ref0b);
+                       if (is_var) {
+                               if (_ref0 == r.ref) putchar("ACGTN"[r.alt]);
+                               else printf("%c,%c", "ACGTN"[r.ref], "ACGTN"[r.alt]);
+                       } else putchar('.');
+                       printf("\t%d\t", qref);
+                       if (!tot) printf("Q13\t");
+                       else printf(".\t");
+                       for (i = depth = 0, sqr_sum = 0; i < n; ++i) {
+                               depth += n_plp[i];
+                               for (j = 0; j < n_plp[i]; ++j) {
+                                       int q = plp[i][j].b->core.qual;
+                                       if (q > conf->max_mq) q = conf->max_mq;
+                                       sqr_sum += q * q;
+                               }
+                       }
+                       rms_q = (int)(sqrt((double)sqr_sum / depth) + .499);
+                       printf("DP=%d;MQ=%d", depth, rms_q);
+                       if (tot) {
+                               printf(";AF=%.3lf", 1. - r.f_em);
+                               if (level >= 2) printf(";AFE=%.3lf", 1-r.f_exp);
+                               if (conf->flag & MPLP_AFALL) {
+                                       printf(";AF0=%.3lf;AFN=%.3lf", 1-r.f_naive, 1-r.f_nielsen);
+                                       if (conf->flag & MPLP_AFS) printf(";AFB=%.3lf", 1-r.f_map);
+                               }
+                       }
+                       printf("\tGT:GQ:DP");
+                       if (tot) {
+                               for (i = 0; i < n; ++i) {
+                                       int x = mc_call_gt(ma, r.f_exp, i);
+                                       printf("\t%c/%c:%d:%d", "10"[((x&3)==2)], "10"[((x&3)>0)], x>>2, n_plp[i]);
+                               }
+                       } else for (i = 0; i < n; ++i) printf("\t./.:0:0");
+                       putchar('\n');
+                       if (N % MPLP_AFS_BLOCK == 0) mc_dump_afs(ma);
+               } else {
+                       printf("%s\t%d\t%c", h->target_name[tid], pos + 1, (ref && pos < ref_len)? ref[pos] : 'N');
+                       for (i = 0; i < n; ++i) {
+                               int j;
+                               printf("\t%d\t", n_plp[i]);
+                               if (n_plp[i] == 0) printf("*\t*");
+                               else {
+                                       for (j = 0; j < n_plp[i]; ++j)
+                                               pileup_seq(plp[i] + j, pos, ref_len, ref);
+                                       putchar('\t');
+                                       for (j = 0; j < n_plp[i]; ++j) {
+                                               const bam_pileup1_t *p = plp[i] + j;
+                                               int c = bam1_qual(p->b)[p->qpos] + 33;
+                                               if (c > 126) c = 126;
+                                               putchar(c);
+                                       }
+                               }
+                       }
+                       putchar('\n');
+               }
+       }
+       if ((conf->flag&MPLP_VCF) && (conf->flag&MPLP_AFS)) mc_dump_afs(ma);
+       if (hash) { // free the hash table
+               khint_t k;
+               for (k = kh_begin(hash); k < kh_end(hash); ++k)
+                       if (kh_exist(hash, k)) free(kh_val(hash, k));
+               kh_destroy(64, hash);
+       }
+       mc_destroy(ma);
+       bam_mplp_destroy(iter);
+       bam_header_destroy(h);
+       for (i = 0; i < n; ++i) {
+               bam_close(data[i]->fp);
+               if (data[i]->iter) bam_iter_destroy(data[i]->iter);
+               free(data[i]);
+       }
+       free(data); free(plp); free(ref); free(n_plp);
+       return 0;
+}
+
+int bam_mpileup(int argc, char *argv[])
+{
+       int c;
+       mplp_conf_t mplp;
+       memset(&mplp, 0, sizeof(mplp_conf_t));
+       mplp.max_mq = 60;
+       mplp.prior_type = MC_PTYPE_FULL;
+       mplp.theta = 1e-3;
+       while ((c = getopt(argc, argv, "vVcF2Sf:r:l:VM:q:t:")) >= 0) {
+               switch (c) {
+               case 't': mplp.theta = atof(optarg); break;
+               case '2': mplp.prior_type = MC_PTYPE_COND2; break;
+               case 'f':
+                       mplp.fai = fai_load(optarg);
+                       if (mplp.fai == 0) return 1;
+                       break;
+               case 'r': mplp.reg = strdup(optarg); break;
+               case 'l': mplp.fn_pos = strdup(optarg); break;
+               case 'V':
+               case 'c': mplp.flag |= MPLP_VCF; break;
+               case 'S': mplp.flag |= MPLP_AFS; break;
+               case 'F': mplp.flag |= MPLP_AFALL; break;
+               case 'v': mplp.flag |= MPLP_VAR; break;
+               case 'M': mplp.max_mq = atoi(optarg); break;
+               case 'q': mplp.min_mq = atoi(optarg); break;
+               }
+       }
+       if (argc == 1) {
+               fprintf(stderr, "\n");
+               fprintf(stderr, "Usage:   samtools mpileup [options] in1.bam [in2.bam [...]]\n\n");
+               fprintf(stderr, "Options: -f FILE     reference sequence file [null]\n");
+               fprintf(stderr, "         -r STR      region in which pileup is generated [null]\n");
+               fprintf(stderr, "         -l FILE     list of positions (format: chr pos) [null]\n");
+               fprintf(stderr, "         -M INT      cap mapping quality at INT [%d]\n", mplp.max_mq);
+               fprintf(stderr, "         -q INT      filter out alignment with MQ smaller than INT [%d]\n", mplp.min_mq);
+               fprintf(stderr, "         -t FLOAT    scaled mutation rate [%lg]\n", mplp.theta);
+               fprintf(stderr, "         -c          generate VCF output (consensus calling)\n");
+               fprintf(stderr, "         -v          show variant sites only\n");
+               fprintf(stderr, "         -S          calculate AFS (slow, to stderr)\n");
+               fprintf(stderr, "         -2          conditional prior\n");
+               fprintf(stderr, "\n");
+               fprintf(stderr, "Notes: Assuming error independency and diploid individuals.\n\n");
+               return 1;
+       }
+       mpileup(&mplp, argc - optind, argv + optind);
+       free(mplp.reg);
+       if (mplp.fai) fai_destroy(mplp.fai);
+       return 0;
+}