def main(reads_file, ref_file, dir, error_rate): sys.stderr.write("Reading reference" + "\n") ref = sorted(list(SeqIO.parse_fasta(open(ref_file, "r"))), key=lambda rec: len(rec))[-1] ref = Contig(ref.seq, ref.id) refs = ContigCollection() for i in range(0, len(ref) - 500, 500): if random.random() > 0.95: tmp = list(ref.segment(i, i + 500).Seq()) for j in range(error_rate * 500 / 100): pos = random.randint(0, 499) tmp[pos] = basic.rc[tmp[pos]] refs.add( Contig("".join(tmp), ref.id + "(" + str(i) + "," + str(i + 500) + ")")) refs.print_names(sys.stderr) sys.stderr.write("Reading reads" + "\n") reads = ReadCollection() reads.loadFromFasta(open(reads_file, "r")) sys.stderr.write("Aligning reads" + "\n") basic.ensure_dir_existance(dir) aligner = Aligner(DirDistributor(dir)) aligner.alignReadCollection(reads, refs) sys.stderr.write("Analysing alignments" + "\n") alignments = [] for read in reads: alignments.extend(read.alignments) alignments = filter(lambda al: len(al) > 450, alignments) alignments = sorted(alignments, key=lambda al: (al.seg_to.contig.id, al.seg_from.contig.id)) scorer = Scorer() scorer.scores.homo_score = 3 scorer.scores.ins_score = 5 scorer.scores.del_score = 5 cnt = 0 for contig, iter in itertools.groupby(alignments, key=lambda al: al.seg_to.contig): iter = list(iter) sys.stderr.write(str(contig) + " " + str(len(iter)) + "\n") if len(iter) < 150: for al in iter: print scorer.accurateScore(al.matchingSequence(), params.alignment_correction_radius) cnt += 1 if cnt >= 5000: break if cnt >= 5000: break
def main(contigs_file, contig_name, reads_file, dir, k, initial_reads1, initial_reads2): basic.ensure_dir_existance(dir) basic.CreateLog(dir) dd = DirDistributor(os.path.join(dir, "alignments")) aligner = Aligner(dd) contigs = ContigStorage().loadFromFasta(open(contigs_file, "r"), False) # contig = contigs[contig_name].asSegment().prefix(length=2000).asContig() contig = contigs[contig_name] reads = ContigStorage().loadFromFasta(open(reads_file, "r"), False) reads1 = ContigStorage() reads2 = ContigStorage() cnt = 0 for read in reads.unique(): cnt += 1 # if cnt % 2 == 0: if read.id in initial_reads1: reads1.add(read) elif read.id in initial_reads2: reads2.add(read) polisher = Polisher(aligner, dd) contig1 = contig contig2 = contig scorer = Scorer() for i in range(3): diff = 0 print "Iteration", i als1 = fixAlDir(aligner.overlapAlign(reads1.unique(), ContigStorage([contig])), contig) als2 = fixAlDir(aligner.overlapAlign(reads2.unique(), ContigStorage([contig])), contig) contig1 = Contig(polisher.polishSmallSegment(contig.asSegment(), als1).seg_from.Seq(), "1") contig2 = Contig(polisher.polishSmallSegment(contig.asSegment(), als2).seg_from.Seq(), "2") al = aligner.overlapAlign([contig1], ContigStorage([contig2])).next() als1 = fixAlDir(aligner.overlapAlign(reads.unique(), ContigStorage([contig1])), contig1) als1 = filter(lambda al: len(al.seg_to) > len(al.seg_to.contig) - 100, als1) als2 = fixAlDir(aligner.overlapAlign(reads.unique(), ContigStorage([contig2])), contig2) als2 = filter(lambda al: len(al.seg_to) > len(al.seg_to.contig) - 100, als2) als1 = sorted(als1, key = lambda al: al.seg_from.contig.id) als2 = sorted(als2, key = lambda al: al.seg_from.contig.id) reads1 = ContigStorage() reads2 = ContigStorage() dp = scorer.accurateScore(al.matchingSequence(), 10) #1 - al.percentIdentity() als_map = dict() for al in als1: als_map[al.seg_from.contig.id] = [al] for al in als2: if al.seg_from.contig.id in als_map: als_map[al.seg_from.contig.id].append(al) com_res = [] diffs = [] for tmp_als in als_map.values(): if len(tmp_als) != 2: continue al1 = tmp_als[0] al2 = tmp_als[1] print al1, al2 assert al1.seg_from.contig == al2.seg_from.contig pi1 = scorer.accurateScore(al1.matchingSequence(), 10) # al1.percentIdentity() pi2 = scorer.accurateScore(al2.matchingSequence(), 10) # al2.percentIdentity() com_res.append((al1, al2, pi1 - pi2)) diffs.append(pi1 - pi2) diffs = sorted(diffs) th1 = diffs[len(diffs) / 4] th2 = diffs[len(diffs) * 3 / 4] print "Thresholds:", th1, th2 for al1, al2, diff in com_res: if diff < th1: reads1.add(al1.seg_from.contig) elif diff > th2: reads2.add(al2.seg_from.contig) # if pi1 > pi2 + dp / 4: # reads1.add(al1.seg_from.contig) # elif pi2 > pi1 + dp / 4: # reads2.add(al2.seg_from.contig) # diff += abs(pi1 - pi2) print float(diff) / len(als1), len(reads1) / 2, len(reads2) / 2 al = aligner.overlapAlign([contig1], ContigStorage([contig2])).next() print al print "\n".join(al.asMatchingStrings2()) for read in reads1: if read.id in initial_reads1: sys.stdout.write(read.id + " ") print "" for read in reads2: if read.id in initial_reads2: sys.stdout.write(read.id + " ") print "" contig1 = prolong(aligner, polisher, contig1, reads1) contig2 = prolong(aligner, polisher, contig2, reads2) contig1.id = "1" contig2.id = "2" out = open(os.path.join(dir, "copies.fasta"), "w") SeqIO.write(contig1, out, "fasta") SeqIO.write(contig2, out, "fasta") out.close() out = open(os.path.join(dir, "reads1.fasta"), "w") for read in reads1.unique(): SeqIO.write(read, out, "fasta") out.close() out = open(os.path.join(dir, "reads2.fasta"), "w") for read in reads2.unique(): SeqIO.write(read, out, "fasta") out.close() print "Finished"