#!/usr/bin/env k8 var getopt = function(args, ostr) { var oli; // option letter list index if (typeof(getopt.place) == 'undefined') getopt.ind = 0, getopt.arg = null, getopt.place = -1; if (getopt.place == -1) { // update scanning pointer if (getopt.ind >= args.length || args[getopt.ind].charAt(getopt.place = 0) != '-') { getopt.place = -1; return null; } if (getopt.place + 1 < args[getopt.ind].length && args[getopt.ind].charAt(++getopt.place) == '-') { // found "--" ++getopt.ind; getopt.place = -1; return null; } } var optopt = args[getopt.ind].charAt(getopt.place++); // character checked for validity if (optopt == ':' || (oli = ostr.indexOf(optopt)) < 0) { if (optopt == '-') return null; // if the user didn't specify '-' as an option, assume it means null. if (getopt.place < 0) ++getopt.ind; return '?'; } if (oli+1 >= ostr.length || ostr.charAt(++oli) != ':') { // don't need argument getopt.arg = null; if (getopt.place < 0 || getopt.place >= args[getopt.ind].length) ++getopt.ind, getopt.place = -1; } else { // need an argument if (getopt.place >= 0 && getopt.place < args[getopt.ind].length) getopt.arg = args[getopt.ind].substr(getopt.place); else if (args.length <= ++getopt.ind) { // no arg getopt.place = -1; if (ostr.length > 0 && ostr.charAt(0) == ':') return ':'; return '?'; } else getopt.arg = args[getopt.ind]; // white space getopt.place = -1; ++getopt.ind; } return optopt; } var re_cs = /([:=*+-])(\d+|[A-Za-z]+)/g; var c, min_cov_len = 10000, min_var_len = 50000, gap_thres = 50, min_mapq = 5; while ((c = getopt(arguments, "l:L:g:q:")) != null) { if (c == 'l') min_cov_len = parseInt(getopt.arg); else if (c == 'L') min_var_len = parseInt(getopt.arg); else if (c == 'g') gap_thres = parseInt(getopt.arg); else if (c == 'q') min_mapq = parseInt(getopt.arg); } if (arguments.length == getopt.ind) { print("Usage: k8 paf2diff.js [options] "); print("Options:"); print(" -l INT min alignment length to compute coverage ["+min_cov_len+"]"); print(" -L INT min alignment length to call variants ["+min_var_len+"]"); print(" -q INT min mapping quality ["+min_mapq+"]"); print(" -g INT short/long gap threshold (for statistics only) ["+gap_thres+"]"); exit(1); } var file = arguments[getopt.ind] == '-'? new File() : new File(arguments[getopt.ind]); var buf = new Bytes(); var tot_len = 0, n_sub = [0, 0, 0], n_ins = [0, 0, 0, 0], n_del = [0, 0, 0, 0]; function count_var(o) { if (o[3] > 1) return; if (o[5] == '-' && o[6] == '-') return; if (o[5] == '-') { // insertion var l = o[6].length; if (l == 1) ++n_ins[0]; else if (l == 2) ++n_ins[1]; else if (l < gap_thres) ++n_ins[2]; else ++n_ins[3]; } else if (o[6] == '-') { // deletion var l = o[5].length; if (l == 1) ++n_del[0]; else if (l == 2) ++n_del[1]; else if (l < gap_thres) ++n_del[2]; else ++n_del[3]; } else { ++n_sub[0]; var s = o[5] + o[6]; if (s == 'ag' || s == 'ga' || s == 'ct' || s == 'tc') ++n_sub[1]; else ++n_sub[2]; } } var a = [], out = []; var c1_ctg = null, c1_start = 0, c1_end = 0, c1_counted = false, c1_len = 0; while (file.readline(buf) >= 0) { var line = buf.toString(); if (!/\ts2:i:/.test(line)) continue; // skip secondary alignments var m, t = line.split("\t", 12); for (var i = 6; i <= 11; ++i) t[i] = parseInt(t[i]); if (t[10] < min_cov_len || t[11] < min_mapq) continue; var ctg = t[5], x = t[7], end = t[8]; // compute regions covered by 1 contig if (ctg != c1_ctg || x >= c1_end) { if (c1_counted && c1_end > c1_start) { c1_len += c1_end - c1_start; print('R', c1_ctg, c1_start, c1_end); } c1_ctg = ctg, c1_start = x, c1_end = end; c1_counted = (t[10] >= min_var_len); } else if (end > c1_end) { // overlap if (c1_counted && x > c1_start) { c1_len += x - c1_start; print('R', c1_ctg, c1_start, x); } c1_start = c1_end, c1_end = end; c1_counted = (t[10] >= min_var_len); } else { // contained if (c1_counted && x > c1_start) { c1_len += x - c1_start; print('R', c1_ctg, c1_start, x); } c1_start = end; } // output variants ahead of this alignment while (out.length) { if (out[0][0] != ctg || out[0][2] <= x) { count_var(out[0]); print('V', out[0].join("\t")); out.shift(); } else break; } // update coverage for (var i = 0; i < out.length; ++i) if (out[i][1] >= x && out[i][2] <= end) ++out[i][3]; // drop alignments that don't overlap with the current one var k = 0; for (var i = 0; i < a.length; ++i) if (a[0][0] == ctg && a[0][2] > x) a[k++] = a[i]; a.length = k; // core loop if (t[10] >= min_var_len) { if ((m = /\tcs:Z:(\S+)/.exec(line)) == null) continue; // no cs tag var cs = m[1]; var blen = 0, n_diff = 0; tot_len += t[10]; while ((m = re_cs.exec(cs)) != null) { var cov = 1; if (m[1] == '*' || m[1] == '+' || m[1] == '-') for (var i = 0; i < a.length; ++i) if (a[0][2] > x) ++cov; if (m[1] == '=' || m[1] == ':') { var l = m[1] == '='? m[2].length : parseInt(m[2]); x += l, blen += l; } else if (m[1] == '*') { out.push([t[5], x, x+1, cov, t[11], m[2].charAt(0), m[2].charAt(1)]); ++x, ++blen, ++n_diff; } else if (m[1] == '+') { out.push([t[5], x, x, cov, t[11], '-', m[2]]); ++blen, ++n_diff; } else if (m[1] == '-') { out.push([t[5], x, x + m[2].length, cov, t[11], m[2], '-']); x += m[2].length, ++blen, ++n_diff; } } } a.push([t[5], t[7], t[8]]); } if (c1_counted && c1_end > c1_start) { c1_len += c1_end - c1_start; print('R', c1_ctg, c1_start, c1_end); } while (out.length) { count_var(out[0]); print('V', out[0].join("\t")); out.shift(); } //warn(tot_len + " alignment columns considered in calling"); warn(c1_len + " reference bases covered by exactly one contig"); warn(n_sub[0] + " substitutions; ts/tv = " + (n_sub[1]/n_sub[2]).toFixed(3)); warn(n_del[0] + " 1bp deletions"); warn(n_ins[0] + " 1bp insertions"); warn(n_del[1] + " 2bp deletions"); warn(n_ins[1] + " 2bp insertions"); warn(n_del[2] + " [3,"+gap_thres+") deletions"); warn(n_ins[2] + " [3,"+gap_thres+") insertions"); warn(n_del[3] + " >="+gap_thres+" deletions"); warn(n_ins[3] + " >="+gap_thres+" insertions"); buf.destroy(); file.close();