sam_view.c

/*  sam_view.c -- SAM<->BAM<->CRAM conversion.

    Copyright (C) 2009-2019 Genome Research Ltd.
    Portions copyright (C) 2009, 2011, 2012 Broad Institute.

    Author: Heng Li <lh3@sanger.ac.uk>

Permission is hereby granted, free of charge, to any person obtaining a copy
of this software and associated documentation files (the "Software"), to deal
in the Software without restriction, including without limitation the rights
to use, copy, modify, merge, publish, distribute, sublicense, and/or sell
copies of the Software, and to permit persons to whom the Software is
furnished to do so, subject to the following conditions:

The above copyright notices and this permission notice shall be included in
all copies or substantial portions of the Software.

THE SOFTWARE IS PROVIDED "AS IS", WITHOUT WARRANTY OF ANY KIND, EXPRESS OR
IMPLIED, INCLUDING BUT NOT LIMITED TO THE WARRANTIES OF MERCHANTABILITY,
FITNESS FOR A PARTICULAR PURPOSE AND NONINFRINGEMENT. IN NO EVENT SHALL
THE AUTHORS OR COPYRIGHT HOLDERS BE LIABLE FOR ANY CLAIM, DAMAGES OR OTHER
LIABILITY, WHETHER IN AN ACTION OF CONTRACT, TORT OR OTHERWISE, ARISING
FROM, OUT OF OR IN CONNECTION WITH THE SOFTWARE OR THE USE OR OTHER
DEALINGS IN THE SOFTWARE.  */

#include <config.h>

#include <stdlib.h>
#include <string.h>
#include <strings.h>
#include <stdio.h>
#include <unistd.h>
#include <math.h>
#include <inttypes.h>
#include <getopt.h>
#include "htslib/sam.h"
#include "htslib/faidx.h"
#include "htslib/khash.h"
#include "htslib/thread_pool.h"
#include "samtools.h"
#include "sam_opts.h"
#include "bedidx.h"

KHASH_SET_INIT_STR(rg)
KHASH_SET_INIT_STR(tv)

typedef khash_t(rg) *rghash_t;
typedef khash_t(tv) *tvhash_t;

// This structure contains the settings for a samview run
typedef struct samview_settings {
    rghash_t rghash;
    tvhash_t tvhash;
    int min_mapQ;
    int flag_on;
    int flag_off;
    int flag_alloff;
    int min_qlen;
    int remove_B;
    uint32_t subsam_seed;
    double subsam_frac;
    char* library;
    void* bed;
    size_t remove_aux_len;
    char** remove_aux;
    int multi_region;
    char* tag;
} samview_settings_t;


// TODO Add declarations of these to a viable htslib or samtools header
extern const char *bam_get_library(sam_hdr_t *header, const bam1_t *b);
extern int bam_remove_B(bam1_t *b);

// Returns 0 to indicate read should be output 1 otherwise
static int process_aln(const sam_hdr_t *h, bam1_t *b, samview_settings_t* settings)
{
    if (settings->remove_B) bam_remove_B(b);
    if (settings->min_qlen > 0) {
        int k, qlen = 0;
        uint32_t *cigar = bam_get_cigar(b);
        for (k = 0; k < b->core.n_cigar; ++k)
            if ((bam_cigar_type(bam_cigar_op(cigar[k]))&1) || bam_cigar_op(cigar[k]) == BAM_CHARD_CLIP)
                qlen += bam_cigar_oplen(cigar[k]);
        if (qlen < settings->min_qlen) return 1;
    }
    if (b->core.qual < settings->min_mapQ || ((b->core.flag & settings->flag_on) != settings->flag_on) || (b->core.flag & settings->flag_off))
        return 1;
    if (settings->flag_alloff && ((b->core.flag & settings->flag_alloff) == settings->flag_alloff))
        return 1;
    if (!settings->multi_region && settings->bed && (b->core.tid < 0 || !bed_overlap(settings->bed, sam_hdr_tid2name(h, b->core.tid), b->core.pos, bam_endpos(b))))
        return 1;
    if (settings->subsam_frac > 0.) {
        uint32_t k = __ac_Wang_hash(__ac_X31_hash_string(bam_get_qname(b)) ^ settings->subsam_seed);
        if ((double)(k&0xffffff) / 0x1000000 >= settings->subsam_frac) return 1;
    }
    if (settings->rghash) {
        uint8_t *s = bam_aux_get(b, "RG");
        if (s) {
            khint_t k = kh_get(rg, settings->rghash, (char*)(s + 1));
            if (k == kh_end(settings->rghash)) return 1;
        }
    }
    if (settings->tvhash && settings->tag) {
        uint8_t *s = bam_aux_get(b, settings->tag);
        if (s) {
            khint_t k = kh_get(tv, settings->tvhash, (char*)(s + 1));
            if (k == kh_end(settings->tvhash)) return 1;
        } else {
            return 1;
        }
    }
    if (settings->library) {
        const char *p = bam_get_library((sam_hdr_t*)h, b);
        if (!p || strcmp(p, settings->library) != 0) return 1;
    }
    if (settings->remove_aux_len) {
        size_t i;
        for (i = 0; i < settings->remove_aux_len; ++i) {
            uint8_t *s = bam_aux_get(b, settings->remove_aux[i]);
            if (s) {
                bam_aux_del(b, s);
            }
        }
    }
    return 0;
}

static int usage(FILE *fp, int exit_status, int is_long_help);

static int add_read_group_single(const char *subcmd, samview_settings_t *settings, char *name)
{
    char *d = strdup(name);
    int ret = 0;

    if (d == NULL) goto err;

    if (settings->rghash == NULL) {
        settings->rghash = kh_init(rg);
        if (settings->rghash == NULL) goto err;
    }

    kh_put(rg, settings->rghash, d, &ret);
    if (ret == -1) goto err;
    if (ret ==  0) free(d); /* Duplicate */
    return 0;

 err:
    print_error(subcmd, "Couldn't add \"%s\" to read group list: memory exhausted?", name);
    free(d);
    return -1;
}

static int add_read_groups_file(const char *subcmd, samview_settings_t *settings, char *fn)
{
    FILE *fp;
    char buf[1024];
    int ret = 0;
    if (settings->rghash == NULL) {
        settings->rghash = kh_init(rg);
        if (settings->rghash == NULL) {
            perror(NULL);
            return -1;
        }
    }

    fp = fopen(fn, "r");
    if (fp == NULL) {
        print_error_errno(subcmd, "failed to open \"%s\" for reading", fn);
        return -1;
    }

    while (ret != -1 && !feof(fp) && fscanf(fp, "%1023s", buf) > 0) {
        char *d = strdup(buf);
        if (d != NULL) {
            kh_put(rg, settings->rghash, d, &ret);
            if (ret == 0) free(d); /* Duplicate */
        } else {
            ret = -1;
        }
    }
    if (ferror(fp)) ret = -1;
    if (ret == -1) {
        print_error_errno(subcmd, "failed to read \"%s\"", fn);
    }
    fclose(fp);
    return (ret != -1) ? 0 : -1;
}

static int add_tag_value_single(const char *subcmd, samview_settings_t *settings, char *name)
{
    char *d = strdup(name);
    int ret = 0;

    if (d == NULL) goto err;

    if (settings->tvhash == NULL) {
        settings->tvhash = kh_init(tv);
        if (settings->tvhash == NULL) goto err;
    }

    kh_put(tv, settings->tvhash, d, &ret);
    if (ret == -1) goto err;
    if (ret ==  0) free(d); /* Duplicate */
    return 0;

 err:
    print_error(subcmd, "Couldn't add \"%s\" to tag values list: memory exhausted?", name);
    free(d);
    return -1;
}

static int add_tag_values_file(const char *subcmd, samview_settings_t *settings, char *fn)
{
    FILE *fp;
    char buf[1024];
    int ret = 0;
    if (settings->tvhash == NULL) {
        settings->tvhash = kh_init(tv);
        if (settings->tvhash == NULL) {
            perror(NULL);
            return -1;
        }
    }

    fp = fopen(fn, "r");
    if (fp == NULL) {
        print_error_errno(subcmd, "failed to open \"%s\" for reading", fn);
        return -1;
    }

    while (ret != -1 && !feof(fp) && fscanf(fp, "%1023s", buf) > 0) {
        char *d = strdup(buf);
        if (d != NULL) {
            kh_put(tv, settings->tvhash, d, &ret);
            if (ret == 0) free(d); /* Duplicate */
        } else {
            ret = -1;
        }
    }
    if (ferror(fp)) ret = -1;
    if (ret == -1) {
        print_error_errno(subcmd, "failed to read \"%s\"", fn);
    }
    fclose(fp);
    return (ret != -1) ? 0 : -1;
}

static inline int check_sam_write1(samFile *fp, const sam_hdr_t *h, const bam1_t *b, const char *fname, int *retp)
{
    int r = sam_write1(fp, h, b);
    if (r >= 0) return r;

    if (fname) print_error_errno("view", "writing to \"%s\" failed", fname);
    else print_error_errno("view", "writing to standard output failed");

    *retp = EXIT_FAILURE;
    return r;
}

int main_samview(int argc, char *argv[])
{
    int c, is_header = 0, is_header_only = 0, ret = 0, compress_level = -1, is_count = 0, has_index_file = 0, no_pg = 0;
    int64_t count = 0;
    samFile *in = 0, *out = 0, *un_out=0;
    FILE *fp_out = NULL;
    sam_hdr_t *header = NULL;
    char out_mode[5], out_un_mode[5], *out_format = "";
    char *fn_in = 0, *fn_idx_in = 0, *fn_out = 0, *fn_fai = 0, *q, *fn_un_out = 0;
    char *fn_out_idx = NULL, *fn_un_out_idx = NULL, *arg_list = NULL;
    sam_global_args ga = SAM_GLOBAL_ARGS_INIT;
    htsThreadPool p = {NULL, 0};
    int filter_state = ALL, filter_op = 0;
    int result;

    samview_settings_t settings = {
        .rghash = NULL,
        .tvhash = NULL,
        .min_mapQ = 0,
        .flag_on = 0,
        .flag_off = 0,
        .flag_alloff = 0,
        .min_qlen = 0,
        .remove_B = 0,
        .subsam_seed = 0,
        .subsam_frac = -1.,
        .library = NULL,
        .bed = NULL,
        .multi_region = 0,
        .tag = NULL
    };

    static const struct option lopts[] = {
        SAM_OPT_GLOBAL_OPTIONS('-', 0, 'O', 0, 'T', '@'),
        {"no-PG", no_argument, NULL, 1},
        { NULL, 0, NULL, 0 }
    };

    /* parse command-line options */
    strcpy(out_mode, "w");
    strcpy(out_un_mode, "w");
    if (argc == 1 && isatty(STDIN_FILENO))
        return usage(stdout, EXIT_SUCCESS, 0);

    // Suppress complaints about '?' being an unrecognised option.  Without
    // this we have to put '?' in the options list, which makes it hard to
    // tell a bad long option from the use of '-?' (both return '?' and
    // set optopt to '\0').
    opterr = 0;

    while ((c = getopt_long(argc, argv,
                            "SbBcCt:h1Ho:O:q:f:F:G:ul:r:T:R:d:D:L:s:@:m:x:U:MX",
                            lopts, NULL)) >= 0) {
        switch (c) {
        case 's':
            if ((settings.subsam_seed = strtol(optarg, &q, 10)) != 0) {
                // Convert likely user input 0,1,2,... to pseudo-random
                // values with more entropy and more bits set
                srand(settings.subsam_seed);
                settings.subsam_seed = rand();
            }
            if (q && *q == '.') {
                settings.subsam_frac = strtod(q, &q);
                if (*q) ret = 1;
            } else {
                ret = 1;
            }

            if (ret == 1) {
                print_error("view", "Incorrect sampling argument \"%s\"", optarg);
                goto view_end;
            }
            break;
        case 'm': settings.min_qlen = atoi(optarg); break;
        case 'c': is_count = 1; break;
        case 'S': break;
        case 'b': out_format = "b"; break;
        case 'C': out_format = "c"; break;
        case 't': fn_fai = strdup(optarg); break;
        case 'h': is_header = 1; break;
        case 'H': is_header_only = 1; break;
        case 'o': fn_out = strdup(optarg); break;
        case 'U': fn_un_out = strdup(optarg); break;
        case 'X': has_index_file = 1; break;
        case 'f': settings.flag_on |= strtol(optarg, 0, 0); break;
        case 'F': settings.flag_off |= strtol(optarg, 0, 0); break;
        case 'G': settings.flag_alloff |= strtol(optarg, 0, 0); break;
        case 'q': settings.min_mapQ = atoi(optarg); break;
        case 'u': compress_level = 0; break;
        case '1': compress_level = 1; break;
        case 'l': settings.library = strdup(optarg); break;
        case 'L':
            if ((settings.bed = bed_read(optarg)) == NULL) {
                print_error_errno("view", "Could not read file \"%s\"", optarg);
                ret = 1;
                goto view_end;
            }
            break;
        case 'r':
            if (add_read_group_single("view", &settings, optarg) != 0) {
                ret = 1;
                goto view_end;
            }
            break;
        case 'R':
            if (add_read_groups_file("view", &settings, optarg) != 0) {
                ret = 1;
                goto view_end;
            }
            break;
        case 'd':
            if (strlen(optarg) < 4 || optarg[2] != ':') {
                print_error_errno("view", "Invalid \"tag:value\" option: \"%s\"", optarg);
                ret = 1;
                goto view_end;
            }

            if (settings.tag) {
                if (settings.tag[0] != optarg[0] || settings.tag[1] != optarg[1]) {
                    print_error("view", "Different tag \"%s\" was specified before: \"%s\"", settings.tag, optarg);
                    ret = 1;
                    goto view_end;
                }
            } else {
                if (!(settings.tag = calloc(3, 1))) {
                    print_error("view", "Could not allocate memory for tag: \"%s\"", optarg);
                    ret = 1;
                    goto view_end;
                }
                memcpy(settings.tag, optarg, 2);
            }

            if (add_tag_value_single("view", &settings, optarg+3) != 0) {
                ret = 1;
                goto view_end;
            }
            break;
        case 'D':
            // Allow ";" as delimiter besides ":" to support MinGW CLI POSIX
            // path translation as described at:
            //   http://www.mingw.org/wiki/Posix_path_conversion
            if (strlen(optarg) < 4 || (optarg[2] != ':' && optarg[2] != ';')) {
                print_error_errno("view", "Invalid \"tag:file\" option: \"%s\"", optarg);
                ret = 1;
                goto view_end;
            }

            if (settings.tag) {
                if (settings.tag[0] != optarg[0] || settings.tag[1] != optarg[1]) {
                    print_error("view", "Different tag \"%s\" was specified before: \"%s\"", settings.tag, optarg);
                    ret = 1;
                    goto view_end;
                }
            } else {
                if (!(settings.tag = calloc(3, 1))) {
                    print_error("view", "Could not allocate memory for tag: \"%s\"", optarg);
                    ret = 1;
                    goto view_end;
                }
                memcpy(settings.tag, optarg, 2);
            }

            if (add_tag_values_file("view", &settings, optarg+3) != 0) {
                ret = 1;
                goto view_end;
            }
            break;
                /* REMOVED as htslib doesn't support this
        //case 'x': out_format = "x"; break;
        //case 'X': out_format = "X"; break;
                 */
        case '?':
            if (optopt == '?') {  // '-?' appeared on command line
                return usage(stdout, EXIT_SUCCESS, 1);
            } else {
                if (optopt) { // Bad short option
                    print_error("view", "invalid option -- '%c'", optopt);
                } else { // Bad long option
                    // Do our best.  There is no good solution to finding
                    // out what the bad option was.
                    // See, e.g. https://stackoverflow.com/questions/2723888/where-does-getopt-long-store-an-unrecognized-option
                    if (optind > 0 && strncmp(argv[optind - 1], "--", 2) == 0) {
                        print_error("view", "unrecognised option '%s'",
                                    argv[optind - 1]);
                    }
                }
                return usage(stderr, EXIT_FAILURE, 0);
            }
        case 'B': settings.remove_B = 1; break;
        case 'x':
            {
                if (strlen(optarg) != 2) {
                    fprintf(stderr, "main_samview: Error parsing -x auxiliary tags should be exactly two characters long.\n");
                    return usage(stderr, EXIT_FAILURE, 0);
                }
                settings.remove_aux = (char**)realloc(settings.remove_aux, sizeof(char*) * (++settings.remove_aux_len));
                settings.remove_aux[settings.remove_aux_len-1] = optarg;
            }
            break;
        case 'M': settings.multi_region = 1; break;
        case 1: no_pg = 1; break;
        default:
            if (parse_sam_global_opt(c, optarg, lopts, &ga) != 0)
                return usage(stderr, EXIT_FAILURE, 0);
            break;
        }
    }
    if (fn_fai == 0 && ga.reference) fn_fai = fai_path(ga.reference);
    if (compress_level >= 0 && !*out_format) out_format = "b";
    if (is_header_only) is_header = 1;
    // File format auto-detection first
    if (fn_out)    sam_open_mode(out_mode+1,    fn_out,    NULL);
    if (fn_un_out) sam_open_mode(out_un_mode+1, fn_un_out, NULL);
    // Overridden by manual -b, -C
    if (*out_format)
        out_mode[1] = out_un_mode[1] = *out_format;
    out_mode[2] = out_un_mode[2] = '\0';
    // out_(un_)mode now 1 or 2 bytes long, followed by nul.
    if (compress_level >= 0) {
        char tmp[2];
        tmp[0] = compress_level + '0'; tmp[1] = '\0';
        strcat(out_mode, tmp);
        strcat(out_un_mode, tmp);
    }
    if (argc == optind && isatty(STDIN_FILENO)) {
        print_error("view", "No input provided or missing option argument.");
        return usage(stderr, EXIT_FAILURE, 0); // potential memory leak...
    }

    fn_in = (optind < argc)? argv[optind] : "-";
    if ((in = sam_open_format(fn_in, "r", &ga.in)) == 0) {
        print_error_errno("view", "failed to open \"%s\" for reading", fn_in);
        ret = 1;
        goto view_end;
    }

    if (fn_fai) {
        if (hts_set_fai_filename(in, fn_fai) != 0) {
            fprintf(stderr, "[main_samview] failed to use reference \"%s\".\n", fn_fai);
            ret = 1;
            goto view_end;
        }
    }
    if ((header = sam_hdr_read(in)) == 0) {
        fprintf(stderr, "[main_samview] fail to read the header from \"%s\".\n", fn_in);
        ret = 1;
        goto view_end;
    }
    if (settings.rghash) {
        sam_hdr_remove_lines(header, "RG", "ID", settings.rghash);
    }
    if (!is_count) {
        if ((out = sam_open_format(fn_out? fn_out : "-", out_mode, &ga.out)) == 0) {
            print_error_errno("view", "failed to open \"%s\" for writing", fn_out? fn_out : "standard output");
            ret = 1;
            goto view_end;
        }
        if (fn_fai) {
            if (hts_set_fai_filename(out, fn_fai) != 0) {
                fprintf(stderr, "[main_samview] failed to use reference \"%s\".\n", fn_fai);
                ret = 1;
                goto view_end;
            }
        }

        if (!no_pg) {
            if (!(arg_list = stringify_argv(argc+1, argv-1))) {
                print_error("view", "failed to create arg_list");
                ret = 1;
                goto view_end;
            }
            if (sam_hdr_add_pg(header, "samtools",
                                         "VN", samtools_version(),
                                         arg_list ? "CL": NULL,
                                         arg_list ? arg_list : NULL,
                                         NULL)) {
                print_error("view", "failed to add PG line to the header");
                ret = 1;
                goto view_end;
            }
        }

        if (*out_format || ga.write_index || is_header ||
            out_mode[1] == 'b' || out_mode[1] == 'c' ||
            (ga.out.format != sam && ga.out.format != unknown_format))  {
            if (sam_hdr_write(out, header) != 0) {
                fprintf(stderr, "[main_samview] failed to write the SAM header\n");
                ret = 1;
                goto view_end;
            }
        }
        if (ga.write_index) {
            if (!(fn_out_idx = auto_index(out, fn_out, header))) {
                ret = 1;
                goto view_end;
            }
        }

        if (fn_un_out) {
            if ((un_out = sam_open_format(fn_un_out, out_un_mode, &ga.out)) == 0) {
                print_error_errno("view", "failed to open \"%s\" for writing", fn_un_out);
                ret = 1;
                goto view_end;
            }
            if (fn_fai) {
                if (hts_set_fai_filename(un_out, fn_fai) != 0) {
                    fprintf(stderr, "[main_samview] failed to use reference \"%s\".\n", fn_fai);
                    ret = 1;
                    goto view_end;
                }
            }
            if (*out_format || is_header ||
                out_un_mode[1] == 'b' || out_un_mode[1] == 'c' ||
                (ga.out.format != sam && ga.out.format != unknown_format))  {
                if (sam_hdr_write(un_out, header) != 0) {
                    fprintf(stderr, "[main_samview] failed to write the SAM header\n");
                    ret = 1;
                    goto view_end;
                }
            }
            if (ga.write_index) {
                if (!(fn_un_out_idx = auto_index(un_out, fn_un_out, header))) {
                    ret = 1;
                    goto view_end;
                }
            }
        }
    }
    else {
        if (fn_out) {
            fp_out = fopen(fn_out, "w");
            if (fp_out == NULL) {
                print_error_errno("view", "can't create \"%s\"", fn_out);
                ret = EXIT_FAILURE;
                goto view_end;
            }
        }
    }

    if (ga.nthreads > 1) {
        if (!(p.pool = hts_tpool_init(ga.nthreads))) {
            fprintf(stderr, "Error creating thread pool\n");
            ret = 1;
            goto view_end;
        }
        hts_set_opt(in,  HTS_OPT_THREAD_POOL, &p);
        if (out) hts_set_opt(out, HTS_OPT_THREAD_POOL, &p);
    }
    if (is_header_only) goto view_end; // no need to print alignments

    if (has_index_file) {
        fn_idx_in = (optind+1 < argc)? argv[optind+1] : 0;
        if (fn_idx_in == 0) {
            fprintf(stderr, "[main_samview] incorrect number of arguments for -X option. Aborting.\n");
            return 1;
        }
    }

    if (settings.multi_region) {
        if (!has_index_file && optind < argc - 1) { //regions have been specified in the command line
            settings.bed = bed_hash_regions(settings.bed, argv, optind+1, argc, &filter_op); //insert(1) or filter out(0) the regions from the command line in the same hash table as the bed file
            if (!filter_op)
                filter_state = FILTERED;
        } else if (has_index_file && optind < argc - 2) {
            settings.bed = bed_hash_regions(settings.bed, argv, optind+2, argc, &filter_op); //insert(1) or filter out(0) the regions from the command line in the same hash table as the bed file
            if (!filter_op)
                filter_state = FILTERED;
        } else {
            bed_unify(settings.bed);
        }

        bam1_t *b = bam_init1();
        if (settings.bed == NULL) { // index is unavailable or no regions have been specified
            fprintf(stderr, "[main_samview] no regions or BED file have been provided. Aborting.\n");
        } else {
            hts_idx_t *idx = NULL;
            // If index filename has not been specfied, look in BAM folder
            if (fn_idx_in != 0) {
                idx = sam_index_load2(in, fn_in, fn_idx_in); // load index
            } else {
                idx = sam_index_load(in, fn_in);
            }
            if (idx != NULL) {

                int regcount = 0;

                hts_reglist_t *reglist = bed_reglist(settings.bed, filter_state, &regcount);
                if(reglist) {
                    hts_itr_multi_t *iter = sam_itr_regions(idx, header, reglist, regcount);
                    if (iter) {
                        // fetch alignments
                        while ((result = sam_itr_multi_next(in, iter, b)) >= 0) {
                            if (!process_aln(header, b, &settings)) {
                                if (!is_count) { if (check_sam_write1(out, header, b, fn_out, &ret) < 0) break; }
                                count++;
                            } else {
                                if (un_out) { if (check_sam_write1(un_out, header, b, fn_un_out, &ret) < 0) break; }
                            }
                        }
                        if (result < -1) {
                            fprintf(stderr, "[main_samview] retrieval of region %d failed due to truncated file or corrupt BAM index file\n", iter->curr_tid);
                            ret = 1;
                        }

                        hts_itr_multi_destroy(iter);
                    } else {
                        fprintf(stderr, "[main_samview] iterator could not be created. Aborting.\n");
                    }
                } else {
                    fprintf(stderr, "[main_samview] region list is empty or could not be created. Aborting.\n");
                }
                hts_idx_destroy(idx); // destroy the BAM index
            } else {
                fprintf(stderr, "[main_samview] random alignment retrieval only works for indexed BAM or CRAM files.\n");
            }
        }
        bam_destroy1(b);
    } else {
        if ((has_index_file && optind >= argc - 2) || (!has_index_file && optind >= argc - 1)) { // convert/print the entire file
            bam1_t *b = bam_init1();
            int r;
            while ((r = sam_read1(in, header, b)) >= 0) { // read one alignment from `in'
                if (!process_aln(header, b, &settings)) {
                    if (!is_count) { if (check_sam_write1(out, header, b, fn_out, &ret) < 0) break; }
                    count++;
                } else {
                    if (un_out) { if (check_sam_write1(un_out, header, b, fn_un_out, &ret) < 0) break; }
                }
            }
            if (r < -1) {
                fprintf(stderr, "[main_samview] truncated file.\n");
                ret = 1;
            }
            bam_destroy1(b);
        } else { // retrieve alignments in specified regions
            int i;
            bam1_t *b;
            hts_idx_t *idx = NULL;
            // If index filename has not been specfied, look in BAM folder
            if (fn_idx_in != NULL) {
                idx = sam_index_load2(in, fn_in, fn_idx_in); // load index
            } else {
                idx = sam_index_load(in, fn_in);
            }
            if (idx == 0) { // index is unavailable
                fprintf(stderr, "[main_samview] random alignment retrieval only works for indexed BAM or CRAM files.\n");
                ret = 1;
                goto view_end;
            }
            b = bam_init1();

            for (i = (has_index_file)? optind+2 : optind+1; i < argc; ++i) {
                int result;
                hts_itr_t *iter = sam_itr_querys(idx, header, argv[i]); // parse a region in the format like `chr2:100-200'
                if (iter == NULL) { // region invalid or reference name not found
                    fprintf(stderr, "[main_samview] region \"%s\" specifies an invalid region or unknown reference. Continue anyway.\n", argv[i]);
                    continue;
                }
                // fetch alignments
                while ((result = sam_itr_next(in, iter, b)) >= 0) {
                    if (!process_aln(header, b, &settings)) {
                        if (!is_count) { if (check_sam_write1(out, header, b, fn_out, &ret) < 0) break; }
                        count++;
                    } else {
                        if (un_out) { if (check_sam_write1(un_out, header, b, fn_un_out, &ret) < 0) break; }
                    }
                }
                hts_itr_destroy(iter);
                if (result < -1) {
                    fprintf(stderr, "[main_samview] retrieval of region \"%s\" failed due to truncated file or corrupt BAM index file\n", argv[i]);
                    ret = 1;
                    break;
                }
            }
            bam_destroy1(b);
            hts_idx_destroy(idx); // destroy the BAM index
        }
    }

    if (ga.write_index) {
        if (sam_idx_save(out) < 0) {
            print_error_errno("view", "writing index failed");
            ret = 1;
        }
        if (un_out && sam_idx_save(un_out) < 0) {
            print_error_errno("view", "writing index failed");
            ret = 1;
        }
    }

view_end:
    if (is_count && ret == 0) {
        if (fprintf(fn_out? fp_out : stdout, "%" PRId64 "\n", count) < 0) {
            if (fn_out) print_error_errno("view", "writing to \"%s\" failed", fn_out);
            else print_error_errno("view", "writing to standard output failed");
            ret = EXIT_FAILURE;
        }
    }

    // close files, free and return
    if (in) check_sam_close("view", in, fn_in, "standard input", &ret);
    if (out) check_sam_close("view", out, fn_out, "standard output", &ret);
    if (un_out) check_sam_close("view", un_out, fn_un_out, "file", &ret);
    if (fp_out) fclose(fp_out);

    free(fn_fai); free(fn_out); free(settings.library);  free(fn_un_out);
    sam_global_args_free(&ga);
    if ( header ) sam_hdr_destroy(header);
    if (settings.bed) bed_destroy(settings.bed);
    if (settings.rghash) {
        khint_t k;
        for (k = 0; k < kh_end(settings.rghash); ++k)
            if (kh_exist(settings.rghash, k)) free((char*)kh_key(settings.rghash, k));
        kh_destroy(rg, settings.rghash);
    }
    if (settings.tvhash) {
        khint_t k;
        for (k = 0; k < kh_end(settings.tvhash); ++k)
            if (kh_exist(settings.tvhash, k)) free((char*)kh_key(settings.tvhash, k));
        kh_destroy(tv, settings.tvhash);
    }
    if (settings.remove_aux_len) {
        free(settings.remove_aux);
    }
    if (settings.tag) {
        free(settings.tag);
    }

    if (p.pool)
        hts_tpool_destroy(p.pool);

    if (fn_out_idx)
        free(fn_out_idx);
    if (fn_un_out_idx)
        free(fn_un_out_idx);
    free(arg_list);

    return ret;
}

static int usage(FILE *fp, int exit_status, int is_long_help)
{
    fprintf(fp,
"\n"
"Usage: samtools view [options] <in.bam>|<in.sam>|<in.cram> [region ...]\n"
"\n"
"Options:\n"
// output options
"  -b       output BAM\n"
"  -C       output CRAM (requires -T)\n"
"  -1       use fast BAM compression (implies -b)\n"
"  -u       uncompressed BAM output (implies -b)\n"
"  -h       include header in SAM output\n"
"  -H       print SAM header only (no alignments)\n"
"  -c       print only the count of matching records\n"
"  -o FILE  output file name [stdout]\n"
"  -U FILE  output reads not selected by filters to FILE [null]\n"
// extra input
"  -t FILE  FILE listing reference names and lengths (see long help) [null]\n"
"  -X       include customized index file\n"
// read filters
"  -L FILE  only include reads overlapping this BED FILE [null]\n"
"  -r STR   only include reads in read group STR [null]\n"
"  -R FILE  only include reads with read group listed in FILE [null]\n"
"  -d STR:STR\n"
"           only include reads with tag STR and associated value STR [null]\n"
"  -D STR:FILE\n"
"           only include reads with tag STR and associated values listed in\n"
"           FILE [null]\n"
"  -q INT   only include reads with mapping quality >= INT [0]\n"
"  -l STR   only include reads in library STR [null]\n"
"  -m INT   only include reads with number of CIGAR operations consuming\n"
"           query sequence >= INT [0]\n"
"  -f INT   only include reads with all  of the FLAGs in INT present [0]\n"       //   F&x == x
"  -F INT   only include reads with none of the FLAGS in INT present [0]\n"       //   F&x == 0
"  -G INT   only EXCLUDE reads with all  of the FLAGs in INT present [0]\n"       // !(F&x == x)
"  -s FLOAT subsample reads (given INT.FRAC option value, 0.FRAC is the\n"
"           fraction of templates/read pairs to keep; INT part sets seed)\n"
"  -M       use the multi-region iterator (increases the speed, removes\n"
"           duplicates and outputs the reads as they are ordered in the file)\n"
// read processing
"  -x STR   read tag to strip (repeatable) [null]\n"
"  -B       collapse the backward CIGAR operation\n"
// general options
"  -?       print long help, including note about region specification\n"
"  -S       ignored (input format is auto-detected)\n"
"  --no-PG  do not add a PG line\n");

    sam_global_opt_help(fp, "-.O.T@..");
    fprintf(fp, "\n");

    if (is_long_help)
        fprintf(fp,
"Notes:\n"
"\n"
"1. This command now auto-detects the input format (BAM/CRAM/SAM).\n"
"   Further control over the CRAM format can be specified by using the\n"
"   --output-fmt-option, e.g. to specify the number of sequences per slice\n"
"   and to use avoid reference based compression:\n"
"\n"
"\tsamtools view -C --output-fmt-option seqs_per_slice=5000 \\\n"
"\t   --output-fmt-option no_ref -o out.cram in.bam\n"
"\n"
"   Options can also be specified as a comma separated list within the\n"
"   --output-fmt value too.  For example this is equivalent to the above\n"
"\n"
"\tsamtools view --output-fmt cram,seqs_per_slice=5000,no_ref \\\n"
"\t   -o out.cram in.bam\n"
"\n"
"2. The file supplied with `-t' is SPACE/TAB delimited with the first\n"
"   two fields of each line consisting of the reference name and the\n"
"   corresponding sequence length. The `.fai' file generated by \n"
"   `samtools faidx' is suitable for use as this file. This may be an\n"
"   empty file if reads are unaligned.\n"
"\n"
"3. SAM->BAM conversion:  samtools view -bT ref.fa in.sam.gz\n"
"\n"
"4. BAM->SAM conversion:  samtools view -h in.bam\n"
"\n"
"5. A region should be presented in one of the following formats:\n"
"   `chr1', `chr2:1,000' and `chr3:1000-2,000'. When a region is\n"
"   specified, the input alignment file must be a sorted and indexed\n"
"   alignment (BAM/CRAM) file.\n"
"\n"
"6. Option `-u' is preferred over `-b' when the output is piped to\n"
"   another samtools command.\n"
"\n");

    return exit_status;
}