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new_reads = defaultdict(realign) for r in reads: world = {} sc = 0 for p in reads[r].precursors: world[p] = reads[r].precursors[p].get_score(len(reads[r].sequence)) if sc < world[p]: sc = world[p] new_reads[r] = reads[r] for p ...
def _clean_hits(reads)
Select only best matches
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mode = "r" if bam_fn.endswith("sam") else "rb" handle = pysam.Samfile(bam_fn, mode) reads = defaultdict(realign) for line in handle: chrom = handle.getrname(line.reference_id) # print("%s %s %s %s" % (line.query_name, line.reference_start, line.query_sequence, chrom)) query_...
def _read_bam(bam_fn, precursors)
read bam file and perform realignment of hits
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args = argparse.Namespace() args.fastq = in_fn args.minimum = 1 args.out = op.dirname(in_fn) return collapse_fastq(args)
def _collapse_fastq(in_fn)
collapse reads into unique sequences
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with open(fn) as handle: reads = defaultdict(realign) for line in handle: query_name, seq, chrom, reference_start, end, mism, add = line.split() reference_start = int(reference_start) # chrom = handle.getrname(cols[1]) # print("%s %s %s %s" % (lin...
def _read_pyMatch(fn, precursors)
read pyMatch file and perform realignment of hits
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if subs!="0": subs = [[subs.replace(subs[-2:], ""),subs[-2], subs[-1]]] return subs
def _parse_mut(subs)
Parse mutation tag from miraligner output
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reads = defaultdict(realign) with open(fn) as in_handle: in_handle.next() for line in in_handle: cols = line.strip().split("\t") iso = isomir() query_name, seq = cols[1], cols[0] chrom, reference_start = cols[-2], cols[3] iso.mirna...
def _read_miraligner(fn)
Read ouput of miraligner and create compatible output.
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tool = _get_miraligner() path_db = op.dirname(op.abspath(hairpin)) cmd = "{tool} -freq -i {fn} -o {out_file} -s {species} -db {path_db} -sub 1 -trim 3 -add 3" if not file_exists(out_file): logger.info("Running miraligner with %s" % fn) do.run(cmd.format(**locals()), "miraligner with...
def _cmd_miraligner(fn, out_file, species, hairpin, out)
Run miraligner for miRNA annotation
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args = argparse.Namespace() args.hairpin = hairpin args.sps = species args.gtf = gff3 args.add_extra = True args.files = out_files args.format = "seqbuster" args.out_format = "gff" args.out = out reader(args)
def _mirtop(out_files, hairpin, gff3, species, out)
Convert miraligner to mirtop format
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df = pd.concat(dts) ma = df.pivot(index='isomir', columns='sample', values='counts') ma_mirna = ma ma = ma.fillna(0) ma_mirna['mirna'] = [m.split(":")[0] for m in ma.index.values] ma_mirna = ma_mirna.groupby(['mirna']).sum() ma_mirna = ma_mirna.fillna(0) return ma, ma_mirna
def _merge(dts)
merge multiple samples in one matrix
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ma, ma_mirna = _merge(out_dts) out_ma = op.join(out_dir, "counts.tsv") out_ma_mirna = op.join(out_dir, "counts_mirna.tsv") ma.to_csv(out_ma, sep="\t") ma_mirna.to_csv(out_ma_mirna, sep="\t") return out_ma_mirna, out_ma
def _create_counts(out_dts, out_dir)
Summarize results into single files.
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hairpin, mirna = _download_mirbase(args) precursors = _read_precursor(args.hairpin, args.sps) matures = _read_mature(args.mirna, args.sps) gtf = _read_gtf(args.gtf) out_dts = [] out_files = [] for bam_fn in args.files: sample = op.splitext(op.basename(bam_fn))[0] logger....
def miraligner(args)
Realign BAM hits to miRBAse to get better accuracy and annotation
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cur_dir = os.getcwd() _mkdir(new_dir) os.chdir(new_dir) try: yield finally: os.chdir(cur_dir)
def chdir(new_dir)
stolen from bcbio. Context manager to temporarily change to a new directory. http://lucentbeing.com/blog/context-managers-and-the-with-statement-in-python/
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target = op.join("seqcluster", "flavor") url = "https://github.com/lpantano/seqcluster.git" if not os.path.exists(target): # shutil.rmtree("seqcluster") subprocess.check_call(["git", "clone","-b", "flavor", "--single-branch", url]) return op.abspath(target)
def _get_flavor()
Download flavor from github
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try: from bcbio import install as bcb except: raise ImportError("It needs bcbio to do the quick installation.") path_flavor = _get_flavor() s = {"fabricrc_overrides": {"system_install": path, "local_install": os.path.join(path, "local_install"), ...
def _install(path, args)
small helper for installation in case outside bcbio
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try: from bcbio import install as bcb except: raise ImportError("It needs bcbio to do the quick installation.") bio_data = op.join(path_flavor, "../biodata.yaml") s = {"flavor": path_flavor, # "target": "[brew, conda]", "vm_provider": "novm", "hostname": "...
def _install_data(data_dir, path_flavor, args)
Upgrade required genome data files in place.
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logger.info(args) logger.info("reading sequeces") out_file = os.path.abspath(os.path.splitext(args.json)[0] + "_prediction.json") data = load_data(args.json) out_dir = os.path.abspath(safe_dirs(os.path.join(args.out, "predictions"))) logger.info("make predictions") data = is_tRNA(data...
def predictions(args)
Create predictions of clusters
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# Original Py 2.7 code #data_loci = map(lambda (x): [x, loci[x].chr, int(loci[x].start), int(loci[x].end), loci[x].strand, len(c.loci2seq[x])], c.loci2seq.keys()) # 2to3 suggested Py 3 rewrite data_loci = [[x, loci[x].chr, int(loci[x].start), int(loci[x].end), loci[x].strand, len(c.loci2seq[x])] fo...
def sort_precursor(c, loci)
Sort loci according to number of sequences mapped there.
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data_loci = sort_precursor(clus, loci) current_size = data_loci[0][5] best = 0 for item, locus in enumerate(data_loci): if locus[3] - locus[2] > 70: if locus[5] > current_size * 0.8: best = item break best_loci = data_loci[best] del data_l...
def best_precursor(clus, loci)
Select best precursor asuming size around 100 nt
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_, ext = os.path.splitext(in_file) if ext == ".gz": return gzip.open(in_file, 'rb') if ext in [".fastq", ".fq"]: return open(in_file, 'r') # default to just opening it return open(in_file, "r")
def _open_file(in_file)
From bcbio code
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with open(out, 'w') as out_handle: print(_create_header(mirna, snp, out), file=out_handle, end="") snp_in_mirna = pybedtools.BedTool(snp).intersect(pybedtools.BedTool(mirna), wo=True) for single in snp_in_mirna: if single[10] == "miRNA" and len(single[3]) + len(single[4]) ==...
def select_snps(mirna, snp, out)
Use bedtools to intersect coordinates
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if 1.0 * x/s >= p: return True elif stat.binom_test(x, s, p) > 0.01: return True return False
def up_threshold(x, s, p)
function to decide if similarity is below cutoff
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scores = [] for start in range(0, len(positions) - 17, 5): end = start = 17 scores.add(_enrichment(positions[start:end], positions[:start], positions[end:]))
def _scan(positions)
get the region inside the vector with more expression
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args = _check_args(args) read_stats_file = op.join(args.dir_out, "read_stats.tsv") if file_exists(read_stats_file): os.remove(read_stats_file) bam_file, seq_obj = _clean_alignment(args) logger.info("Parsing matrix file") seqL, y, l = parse_ma_file(seq_obj, args.ffile) # y, l ...
def cluster(args)
Creating clusters
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logger.info("Checking parameters and files") args.dir_out = args.out args.samplename = "pro" global decision_cluster global similar if not os.path.isdir(args.out): logger.warning("the output folder doens't exists") os.mkdirs(args.out) if args.bed and args.gtf: lo...
def _check_args(args)
check arguments before starting analysis.
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total = Counter() if isinstance(seqs, list): if not aligned: l = len([total.update(seqL[s].freq) for s in seqs]) else: l = len([total.update(seqL[s].freq) for s in seqs if seqL[s].align > 0]) elif isinstance(seqs, dict): [total.update(seqs[s].get_freq(seq...
def _total_counts(seqs, seqL, aligned=False)
Counts total seqs after each step
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data_ann_temp = {} data_ann = [] counts = Counter() for lid in c.loci2seq: # original Py 2.7 code #for dbi in loci[lid].db_ann.keys(): # data_ann_temp[dbi] = {dbi: map(lambda (x): loci[lid].db_ann[dbi].ann[x].name, loci[lid].db_ann[dbi].ann.keys())} # suggestion b...
def _get_annotation(c, loci)
get annotation of transcriptional units
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n = len(seqs_freq[seqs_freq.keys()[0]].freq.keys()) y = np.array([0] * n) for s in seqs_freq: x = seqs_freq[s].freq exp = [seqs_freq[s].freq[sam] for sam in samples_order] y = list(np.array(exp) + y) return y
def _sum_by_samples(seqs_freq, samples_order)
Sum sequences of a metacluster by samples.
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logger.info("Creating bed file") bedfile = generate_position_bed(setclus) a = pybedtools.BedTool(bedfile, from_string=True) beds = [] logger.info("Annotating clusters") if hasattr(args, 'list_files'): beds = args.list_files.split(",") for filebed in beds: logger....
def _annotate(args, setclus)
annotate transcriptional units with gtf/bed files provided by -b/g option
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logger.info("Clean bam file with highly repetitive reads with low counts. sum(counts)/n_hits > 1%") bam_file, seq_obj = clean_bam_file(args.afile, args.mask) logger.info("Using %s file" % bam_file) detect_complexity(bam_file, args.ref, args.out) return bam_file, seq_obj
def _clean_alignment(args)
Prepare alignment for cluster detection.
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clus_obj = [] cluster_file = op.join(args.out, "cluster.bed") if not os.path.exists(op.join(args.out, 'list_obj.pk')): if not file_exists(cluster_file): logger.info("Parsing aligned file") logger.info("Merging sequences") bedtools = os.path.join(os.path.dirna...
def _create_clusters(seqL, bam_file, args)
Cluster sequences and create metaclusters with multi-mappers.
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backup = op.join(path, "list_obj_red.pk") if not op.exists(backup): clus_obj = reduceloci(clusL, path) with open(backup, 'wb') as output: pickle.dump(clus_obj, output, pickle.HIGHEST_PROTOCOL) return clus_obj else: logger.info("Loading previous reduced cluste...
def _cleaning(clusL, path)
Load saved cluster and jump to next step
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logger.info("reading sequeces") data = load_data(args.json) logger.info("get sequences from json") #get_sequences_from_cluster() c1, c2 = args.names.split(",") seqs, names = get_sequences_from_cluster(c1, c2, data[0]) loci = get_precursors_from_cluster(c1, c2, data[0]) logger.info("...
def explore(args)
Create mapping of sequences of two clusters
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try: f = open(args.config, 'r') seq_out = open(op.join(args.out, "seqs.fastq"), 'w') ma_out = open(op.join(args.out, "seqs.ma"), 'w') except IOError as e: traceback.print_exc() raise IOError("Can not create output files: %s, %s or read %s" % (op.join(args.out, "seqs....
def prepare(args)
Read all seq.fa files and create a matrix and unique fasta files. The information is :param args: options parsed from command line :param con: logging messages going to console :param log: logging messages going to console and file :returns: files - matrix and fasta files that should be used with ...
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seq_l = {} sample_l = [] idx = 1 for line1 in f: line1 = line1.strip() cols = line1.split("\t") with open(cols[0], 'r') as fasta: sample_l.append(cols[1]) for line in fasta: if line.startswith(">"): idx += 1 ...
def _read_fasta_files(f, args)
read fasta files of each sample and generate a seq_obj with the information of each unique sequence in each sample :param f: file containing the path for each fasta file and the name of the sample. Two column format with `tab` as field separator :returns: * :code:`seq_l`: is a list of seq_obj obje...
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seq_l = {} sample_l = [] idx = 1 p = re.compile("^[ATCGNU]+$") with open(op.join(args.out, "stats_prepare.tsv"), 'w') as out_handle: for line1 in f: line1 = line1.strip() cols = line1.split("\t") # if not is_fastq(cols[0]): # raise Valu...
def _read_fastq_files(f, args)
read fasta files of each sample and generate a seq_obj with the information of each unique sequence in each sample :param f: file containing the path for each fasta file and the name of the sample. Two column format with `tab` as field separator :returns: * :code:`seq_l`: is a list of seq_obj obje...
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skip = 0 if int(min_shared) > len(sample_l): min_shared = len(sample_l) maout.write("id\tseq") for g in sample_l: maout.write("\t%s" % g) for s in seq_l.keys(): seen = sum([1 for g in seq_l[s].group if seq_l[s].group[g] > 0]) if seen < int(min_shared): ...
def _create_matrix_uniq_seq(sample_l, seq_l, maout, out, min_shared)
create matrix counts for each different sequence in all the fasta files :param sample_l: :code:`list_s` is the output of :code:`_read_fasta_files` :param seq_l: :code:`seq_s` is the output of :code:`_read_fasta_files` :param maout: is a file handler to write the matrix count information :param out: is ...
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if not args.bed: raise ValueError("This module needs the bed file output from cluster subcmd.") workdir = op.abspath(op.join(args.out, 'coral')) safe_dirs(workdir) bam_in = op.abspath(args.bam) bed_in = op.abspath(args.bed) reference = op.abspath(args.ref) with chdir(workdir): ...
def run_coral(clus_obj, out_dir, args)
Run some CoRaL modules to predict small RNA function
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ref = os.path.abspath(args.reference) utils.safe_dirs(out_dir) for nc in clus_obj[0]: c = clus_obj[0][nc] loci = c['loci'] out_fa = "cluster_" + nc if loci[0][3] - loci[0][2] < 500: with make_temp_directory() as tmpdir: os.chdir(tmpdir) ...
def is_tRNA(clus_obj, out_dir, args)
Iterates through cluster precursors to predict sRNA types
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score = 0 if os.path.getsize(summary_file) == 0: return 0 with open(summary_file) as in_handle: # header = in_handle.next().strip().split() for line in in_handle: if not line.startswith("--"): pre = line.strip().split() score = pre[-1]...
def _read_tRNA_scan(summary_file)
Parse output from tRNA_Scan
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out_file = fasta_file + "_trnascan" se_file = fasta_file + "_second_str" cmd = "tRNAscan-SE -q -o {out_file} -f {se_file} {fasta_file}" run(cmd.format(**locals())) return out_file, se_file
def _run_tRNA_scan(fasta_file)
Run tRNA-scan-SE to predict tRNA
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multiplier = 1 if mut.startswith("-"): mut = mut[1:] multiplier = -1 nt = mut.strip('0123456789') pos = int(mut[:-2]) * multiplier return nt, pos
def _parse_mut(mut)
Parse mutation field to get position and nts.
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mut = isomir.split(":")[1] if mut == "0": return mut nt, pos = _parse_mut(mut) trim5 = isomir.split(":")[-2] off = -1 * len(trim5) if trim5.islower(): off = len(trim5) if trim5 == "NA" or trim5 == "0": off = 0 # print(isomir) # print([mut, pos, off, nt]) ...
def _get_reference_position(isomir)
Liftover from isomir to reference mature
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pass_pos = [] for isomir in isomirs.iterrows(): mir = isomir[1]["chrom"] mut = isomir[1]["sv"] mut_counts = isomir[1]["counts"] total = mirna.loc[mir, "counts"] * 1.0 - mut_counts mut_diff = isomir[1]["diff"] ratio = mut_counts / total if mut_counts >...
def _get_pct(isomirs, mirna)
Get pct of variants respect to the reference using reads and different sequences
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print("##fileformat=VCFv4.2", file=STDOUT, end="") print("##source=seqbuster2.3", file=STDOUT, end="") print("##reference=mirbase", file=STDOUT, end="") for pos in data: print("##contig=<ID=%s>" % pos["chrom"], file=STDOUT, end="") print('##INFO=<ID=ID,Number=1,Type=String,Description="...
def _print_header(data)
Create vcf header to make a valid vcf.
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id_name = "." qual = "." chrom = data['chrom'] pos = data['pre_pos'] nt_ref = data['nt'][1] nt_snp = data['nt'][0] flt = "PASS" info = "ID=%s" % data['mature'] frmt = "GT:NR:NS" gntp = "%s:%s:%s" % (_genotype(data), data["counts"], data["diff"]) print("\t".join(map(str, ...
def print_vcf(data)
Print vcf line following rules.
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fixed_pos = [] _print_header(pass_pos) for pos in pass_pos: mir = pos["mature"] db_pos = matures[pos["chrom"]] mut = _parse_mut(pos["sv"]) print([db_pos[mir], mut, pos["sv"]]) pos['pre_pos'] = db_pos[mir][0] + mut[1] - 1 pos['nt'] = list(mut[0]) f...
def liftover(pass_pos, matures)
Make position at precursor scale
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global STDOUT isomirs['sv'] = [_get_reference_position(m) for m in isomirs["isomir"]] mirna = isomirs.groupby(['chrom']).sum() sv = isomirs.groupby(['chrom', 'mature', 'sv'], as_index=False).sum() sv["diff"] = isomirs.groupby(['chrom', 'mature', 'sv'], as_index=False).size().reset_index().loc[:...
def create_vcf(isomirs, matures, gtf, vcf_file=None)
Create vcf file of changes for all samples. PASS will be ones with > 3 isomiRs supporting the position and > 30% of reads, otherwise LOW
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fixed_pos = [] for pos in pass_pos: if pos["chrom"] not in gtf: continue db_pos = gtf[pos["chrom"]][0] mut = _parse_mut(pos["sv"]) print([db_pos, pos]) if db_pos[3] == "+": pos['pre_pos'] = db_pos[1] + pos["pre_pos"] + 1 else: ...
def liftover_to_genome(pass_pos, gtf)
Liftover from precursor to genome
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already_in = set() not_in = [] already_in = map(seen.get, seqs) # if isinstance(already_in, list): already_in = filter(None, already_in) not_in = set(seqs) - set(seen.keys()) # for s in seqs: # if s in seen: # already_in.add(seen[s]) # else: # not_in...
def _get_seqs_from_cluster(seqs, seen)
Returns the sequences that are already part of the cluster :param seqs: list of sequences ids :param clus_id: dict of sequences ids that are part of a cluster :returns: * :code:`already_in`list of cluster id that contained some of the sequences * :code:`not_in`list of sequences that don't ...
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filtered = {} n_cluster = 0 large = 0 current = clus_obj.clusid logger.info("Number of loci: %s" % len(clus_obj.loci.keys())) bar = ProgressBar(maxval=len(current)) bar.start() bar.update(0) for itern, idmc in enumerate(current): bar.update(itern) logger.debug("_...
def reduceloci(clus_obj, path)
reduce number of loci a cluster has :param clus_obj: cluster object object :param path: output path
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out_file = op.join(path, 'log', str(idx) + '.bed') with utils.safe_run(out_file): with open(out_file, 'w') as out_handle: for idc in metacluster: for idl in cluster[idc].loci2seq: pos = loci[idl].list() print("\t".join(pos[:4] + [s...
def _write_cluster(metacluster, cluster, loci, idx, path)
For complex meta-clusters, write all the loci for further debug
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global CONFLICT loci = dict(zip(meta, [clusters[idc] for idc in meta])) n_loci = len(meta) n_loci_prev = n_loci + 1 cicle = 0 # [logger.note("BEFORE %s %s %s" % (c.id, idl, len(c.loci2seq[idl]))) for idl in c.loci2seq] internal_cluster = {} if n_loci == 1: n_cluster += 1 ...
def _iter_loci(meta, clusters, s2p, filtered, n_cluster)
Go through all locus and decide if they are part of the same TU or not. :param idx: int cluster id :param s2p: dict with [loci].coverage[start] = # of sequences there :param filtered: dict with clusters object :param n_cluster: int cluster id :return: * filtered: dict of cluster object...
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new_dict = {} n_cluster = 0 logger.debug("_convert_to_cluster: loci %s" % c.loci2seq.keys()) for idl in c.loci2seq: n_cluster += 1 new_c = cluster(n_cluster) #new_c.id_prev = c.id new_c.loci2seq[idl] = c.loci2seq[idl] new_dict[n_cluster] = new_c logger.de...
def _convert_to_clusters(c)
Return 1 cluster per loci
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ma = {} for idc in c: set1 = _get_seqs(c[idc]) [ma.update({(idc, idc2): _common(set1, _get_seqs(c[idc2]), idc, idc2)}) for idc2 in c if idc != idc2 and (idc2, idc) not in ma] # logger.debug("_calculate_similarity_ %s" % ma) return ma
def _calculate_similarity(c)
Get a similarity matrix of % of shared sequence :param c: cluster object :return ma: similarity matrix
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seqs = set() for idl in list_idl.loci2seq: # logger.debug("_get_seqs_: loci %s" % idl) [seqs.add(s) for s in list_idl.loci2seq[idl]] # logger.debug("_get_seqs_: %s" % len(seqs)) return seqs
def _get_seqs(list_idl)
get all sequences in a cluster knowing loci
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c = len(set(s1).intersection(s2)) t = min(len(s1), len(s2)) pct = 1.0 * c / t * t is_gt = up_threshold(pct, t * 1.0, parameters.similar) logger.debug("_common: pct %s of clusters:%s %s = %s" % (1.0 * c / t, i1, i2, is_gt)) if pct < parameters.similar and is_gt and pct > 0: pct = par...
def _common(s1, s2, i1, i2)
calculate the common % percentage of sequences
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1.009011
all_true1 = all([all([common and loci_similarity[(p, c)] > parameters.similar for p in pairs if (p, c) in loci_similarity]) for c in clus_seen]) all_true2 = all([all([common and loci_similarity[(c, p)] > parameters.similar for p in pairs if (c, p) in loci_similarity]) for c in clus_seen]) return all_...
def _is_consistent(pairs, common, clus_seen, loci_similarity)
Check if loci shared that match sequences with all clusters seen until now.
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n_cluster = 0 internal_cluster = {} clus_seen = {} loci_sorted = sorted(loci_similarity.iteritems(), key=operator.itemgetter(1), reverse=True) for pairs, sim in loci_sorted: common = sim > parameters.similar n_cluster += 1 logger.debug("_merge_similar:try new cluster %s"...
def _merge_similar(loci, loci_similarity)
Internal function to reduce loci complexity :param loci: class cluster :param locilen_sorted: list of loci sorted by size :return c: updated class cluster
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logger.debug("_merge_cluster: %s to %s" % (old.id, new.id)) logger.debug("_merge_cluster: add idls %s" % old.loci2seq.keys()) for idl in old.loci2seq: # if idl in new.loci2seq: # new.loci2seq[idl] = list(set(new.loci2seq[idl] + old.loci2seq[idl])) # new.loci2seq[idl] = old.lo...
def _merge_cluster(old, new)
merge one cluster to another
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2.784822
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logger.debug("_solve_conflict: count once") if parameters.decision_cluster == "bayes": return decide_by_bayes(list_c, s2p) loci_similarity = _calculate_similarity(list_c) loci_similarity = sorted(loci_similarity.iteritems(), key=operator.itemgetter(1), reverse=True) common = sum([score ...
def _solve_conflict(list_c, s2p, n_cluster)
Make sure sequences are counts once. Resolve by most-vote or exclussion :params list_c: dict of objects cluster :param s2p: dict of [loci].coverage = # num of seqs :param n_cluster: number of clusters return dict: new set of clusters
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old = c[p[0]] new = c[p[1]] new_c = cluster(n) common = set(_get_seqs(old)).intersection(_get_seqs(new)) for idl in old.loci2seq: in_common = list(set(common).intersection(old.loci2seq[idl])) if len(in_common) > 0: logger.debug("_split_cluster: in_common %s with pair...
def _split_cluster(c, pairs, n)
split cluster by exclussion
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old, new = c[p[0]], c[p[1]] old_size = _get_seqs(old) new_size = _get_seqs(new) logger.debug("_most_vote: size of %s with %s - %s with %s" % (old.id, len(old_size), new.id, len(new_size))) if len(old_size) > len(new_size): keep, remove = old, new else: keep, remove = new, ol...
def _split_cluster_by_most_vote(c, p)
split cluster by most-vote strategy
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global REMOVED init = len(list_c) list_c = {k: v for k, v in list_c.iteritems() if len(_get_seqs(v)) > parameters.min_seqs} logger.debug("_clean_cluster: number of clusters %s " % len(list_c.keys())) list_c = {k: _select_loci(v) for k, v in list_c.iteritems()} end = len(list_c) REMOVED ...
def _clean_cluster(list_c)
Remove cluster with less than 10 sequences and loci with size smaller than 60%
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loci_len = {k: len(v) for k, v in c.loci2seq.iteritems()} logger.debug("_select_loci: number of loci %s" % len(c.loci2seq.keys())) loci_len_sort = sorted(loci_len.iteritems(), key=operator.itemgetter(1), reverse=True) max_size = loci_len_sort[0][1] logger.debug("_select_loci: max size %s" % max...
def _select_loci(c)
Select only loci with most abundant sequences
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first_run = 0 seen_seqs = list() n_cluster += 1 logger.debug("_solve_loci:new cluster %s" % n_cluster) new_c = cluster(n_cluster) for idl, lenl in locilen_sorted: locus_seqs = c.loci2seq[idl] if first_run == 0: seen_seqs = locus_seqs first_run = 1 ...
def _solve_loci_deprecated(c, locilen_sorted, seen_seqs, filtered, maxseq, n_cluster)
internal function to reduce loci complexity The function will read the all loci in a cluster of sequences and will determine if all loci are part of the same transcriptional unit(TU) by most-vote locus or by exclusion of common sequence that are the minority of two loci. :param c: class cluste...
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1.009932
ann = set() if not string: return "This cluster is inter-genic." for item in string: for db in item: ann = ann.union(set(item[db])) return "annotated as: %s ..." % ",".join(list(ann)[:3])
def _get_description(string)
Parse annotation to get nice description
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x = set() for sample in profile: x = x.union(set(profile[sample].keys())) if not x: return '' end, start = max(x), min(x) x = range(start, end, 4) scaled_profile = defaultdict(list) for pos in x: for sample in profile: y = _get_closer(profile[sample],...
def _set_format(profile)
Prepare dict to list of y values with same x
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1.113796
with con: cur = con.cursor() cur.execute("DROP TABLE IF EXISTS clusters;") cur.execute("CREATE TABLE clusters(Id INT, Description TEXT, Locus TEXT, Annotation TEXT, Sequences TEXT, Profile TXT, Precursor TXT)") for c in data[0]: locus = json.dumps(data[0][c]['loci'])...
def _insert_data(con, data)
insert line for each cluster
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loc_id = 1 bedfile_clusters = "" bamfile = pybedtools.BedTool(file_in) bed = pybedtools.BedTool.bam_to_bed(bamfile) for c, start, end, name, q, strand in bed: loc_id += 1 bedfile_clusters += "%s\t%s\t%s\t%s\t%s\t%s\n" % \ (c, start, end, name, loc_id,...
def parse_align_file(file_in)
Parse sam files with aligned sequences
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name = "" index = 1 total = defaultdict(int) with open(in_file) as handle_in: line = handle_in.readline().strip() cols = line.split("\t") samples = cols[2:] for line in handle_in: line = line.strip() cols = line.split("\t") name = ...
def parse_ma_file(seq_obj, in_file)
read seqs.ma file and create dict with sequence object
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3.044256
0.977627
field = field.lower() try: group = cols[2] attrs = cols[8].split(";") name = [attr.strip().split(" ")[1] for attr in attrs if attr.strip().split(" ")[0].lower().endswith(field)] if not name: name = [attr.strip().split(" ")[1] for attr in attrs if attr.strip().spl...
def read_gtf_line(cols, field="name")
parse gtf line to get class/name information
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3.817334
1.010275
strd = "-" if pos_a[2] in pos_b[2]: strd = "+" if pos_a[2] in "+" and pos_b[2] in "+": lento5 = pos_a[0] - pos_b[1] + 1 lento3 = pos_a[1] - pos_b[1] + 1 if pos_a[2] in "+" and pos_b[2] in "-": lento5 = pos_a[1] - pos_b[0] + 1 lento3 = pos_a[0] - pos_b[1] + 1 ...
def _position_in_feature(pos_a, pos_b)
return distance to 3' and 5' end of the feature
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1.537428
1.030674
id_sa, id_ea, id_id, id_idl, id_sta = 1, 2, 3, 4, 5 if type_ann == "bed": id_sb = 7 id_eb = 8 id_stb = 11 id_tag = 9 ida = 0 clus_id = clus_obj.clus loci_id = clus_obj.loci db = os.path.splitext(db)[0] logger.debug("Type:%s\n" % type_ann) for cols in ...
def anncluster(c, clus_obj, db, type_ann, feature_id="name")
intersect transcription position with annotation files
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1.020695
if not genome: logger.info("No genome given. skipping.") return None out_file = op.join(out, op.basename(bam_in) + "_cov.tsv") if file_exists(out_file): return None fai = genome + ".fai" cov = pybedtools.BedTool(bam_in).genome_coverage(g=fai, max=1) cov.saveas(out_fi...
def detect_complexity(bam_in, genome, out)
genome coverage of small RNA
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3.697613
1.015183
seq_obj = defaultdict(int) if mask: mask_file = op.splitext(bam_in)[0] + "_mask.bam" if not file_exists(mask_file): pybedtools.BedTool(bam_file).intersect(b=mask, v=True).saveas(mask_file) bam_in = mask_file out_file = op.splitext(bam_in)[0] + "_rmlw.bam" # bam.i...
def clean_bam_file(bam_in, mask=None)
Remove from alignment reads with low counts and highly # of hits
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2.885152
1.017825
current_loci = {} current_clus = {} # sequence2clusters = [set()] * (max(current_seq.keys()) + 2) sequence2clusters = defaultdict(set) lindex = 0 eindex = 0 previous_id = 0 for line in c.features(): c, start, end, name, score, strand, c_id = line name = int(name.repl...
def detect_clusters(c, current_seq, MIN_SEQ, non_un_gl=False)
Parse the merge file of sequences position to create clusters that will have all sequences that shared any position on the genome :param c: file from bedtools with merge sequence positions :param current_seq: list of sequences :param MIN_SEQ: int cutoff to keep the cluster or not. 10 as default :r...
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3.780112
1.036937
seen = defaultdict(int) metacluster = defaultdict(set) c_index = len(sequence2clusters) logger.info("Creating meta-clusters based on shared sequences: %s" % c_index) meta_idx = 1 bar = ProgressBar(maxval=c_index) bar.start() bar.update() for itern, name in enumerate(sequence2clu...
def _find_metaclusters(clus_obj, sequence2clusters, current_seq, min_seqs)
Mask under same id all clusters that share sequences :param clus_obj: cluster object coming from detect_cluster :param min_seqs: int cutoff to keep the cluster or not. 10 as default :return: updated clus_obj and dict with seq_id: cluster_id
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0.951668
logger.info("Creating meta-clusters based on shared sequences.") seen = defaultdict() metacluster = defaultdict(list) c_index = clus_obj.keys() meta_idx = 0 with ProgressBar(maxval=len(c_index), redirect_stdout=True) as p: for itern, c in enumerate(c_index): p.update(ite...
def _find_families_deprecated(clus_obj, min_seqs)
Mask under same id all clusters that share sequences :param clus_obj: cluster object coming from detect_cluster :param min_seqs: int cutoff to keep the cluster or not. 10 as default :return: updated clus_obj and dict with seq_id: cluster_id
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new_cluster = {} for cid in clus_obj.clus: cluster = clus_obj.clus[cid] cluster.update() logger.debug("peak calling for %s" % cid) bigger = cluster.locimaxid if bigger in clus_obj.loci: s, e = min(clus_obj.loci[bigger].counts.keys()), max(clus_obj.loci[bi...
def peak_calling(clus_obj)
Run peak calling inside each cluster
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if args.fasta: name = None seq = "" reads = dict() with open(args.fasta) as in_handle: for line in in_handle: if line.startswith(">"): if name: reads.update(_generate_reads(seq, name)) se...
def simulate(args)
Main function that manage simulatin of small RNAs
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1.04042
reads = dict() if len(seq) < 130 and len(seq) > 70: reads.update(_mature(seq[:40], 0, name)) reads.update(_mature(seq[-40:], len(seq) - 40, name)) reads.update(_noise(seq, name)) reads.update(_noise(seq, name, 25)) return reads
def _generate_reads(seq, name)
Main function that create reads from precursors
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1.032046
reads = dict() probs = [0.1, 0.2, 0.4, 0.2, 0.1] end = 5 + size error = [-2, -1, 0, 1, 2] for error5 in error: for error3 in error: s = 5 - error5 e = end - error3 seen = subseq[s:e] counts = int(probs[error5 + 2] * probs[error3 + 2] * tot...
def _mature(subseq, absolute, c, size=33, total=5000)
Create mature sequences around start/end
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1.015747
reads = dict() seen = 0 while seen < total: s = random.randint(0, len(seq) - size) e = s + size + random.randint(-5,5) p = random.uniform(0, 0.1) counts = int(p * total) + 1 seen += counts name = "seq_%s_%s_%s_x%s" % (c, s, e, counts) reads[name] ...
def _noise(seq, c, size=33, total=1000)
Create mature sequences around start/end
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out_ma = prefix + ".ma" out_fasta = prefix + ".fasta" out_real = prefix + ".txt" with open(out_ma, 'w') as ma_handle: print("id\tseq\tsample", file=ma_handle, end="") with open(out_fasta, 'w') as fa_handle: with open(out_real, 'w') as read_handle: for idx...
def _write_reads(reads, prefix)
Write fasta file, ma file and real position
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logger.info("Reading sequeces") data = parse_ma_file(args.ma) logger.info("Get sequences from sam") is_align = _read_sam(args.sam) is_json, is_db = _read_json(args.json) res = _summarise_sam(data, is_align, is_json, is_db) _write_suma(res, os.path.join(args.out, "stats_align.dat")) ...
def stats(args)
Create stats from the analysis
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is_json = set() is_db = {} with open(fn_json) as handle: data = json.load(handle) # original Py 2.y core #for item in data[0].values(): # seqs_name = map(lambda (x): x.keys(), item['seqs']) # rewrite by 2to3 for item in list(data[0].values()): ...
def _read_json(fn_json)
read json information
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try: logger.debug(" ".join(str(x) for x in cmd) if not isinstance(cmd, basestring) else cmd) _do_run(cmd, checks, log_stdout) except: if log_error: logger.info("error at command") raise
def run(cmd, data=None, checks=None, region=None, log_error=True, log_stdout=False)
Run the provided command, logging details and checking for errors.
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if isinstance(cmd, basestring): # check for standard or anonymous named pipes if cmd.find(" | ") > 0 or cmd.find(">(") or cmd.find("<("): return "set -o pipefail; " + cmd, True, find_bash() else: return cmd, True, None else: return [str(x) for x in cm...
def _normalize_cmd_args(cmd)
Normalize subprocess arguments to handle list commands, string and pipes. Piped commands set pipefail and require use of bash to help with debugging intermediate errors.
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cmd, shell_arg, executable_arg = _normalize_cmd_args(cmd) s = subprocess.Popen(cmd, shell=shell_arg, executable=executable_arg, stdout=subprocess.PIPE, stderr=subprocess.STDOUT, close_fds=True) debug_stdout = collections.deque(maxlen=100) while 1: ...
def _do_run(cmd, checks, log_stdout=False)
Perform running and check results, raising errors for issues.
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seqs = defaultdict(set) # n = len(list_c.keys()) for c in list_c.values(): for l in c.loci2seq: [seqs[s].add(c.id) for s in c.loci2seq[l]] common = [s for s in seqs if len(seqs[s]) > 1] seqs_in_c = defaultdict(float) for c in list_c.values(): for l in c.loci2seq...
def _dict_seq_locus(list_c, loci_obj, seq_obj)
return dict with sequences = [ cluster1, cluster2 ...]
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for hypo in self.Values(): like = self.Likelihood(data, hypo) self.Mult(hypo, like) self.Normalize()
def Update(self, data)
Updates the PMF with new data. data: string cookie type
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1.0305
mix = self.loci[hypo] like = mix[data] return like
def Likelihood(self, data, hypo)
The likelihood of the data under the hypothesis. data: string cookie type hypo: string bowl ID
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15.598875
1.179912
current = clus_obj.clus clus_seqt = clus_obj.seq clus_locit = clus_obj.loci itern = 0 for idc in current.keys(): itern += 1 timestamp = str(idc) seqListTemp = () f = open("/tmp/"+timestamp+".fa","w") for idl in current[idc].loci2seq.keys(): s...
def show_seq(clus_obj, index)
Get the precursor and map sequences to it. this way we create a positional map.
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1.000623
for ids in s: obj = s[ids] [obj.norm_freq.update({sample: 1.0 * obj.freq[sample] / (t[sample]+1) * 1000000}) for sample in obj.norm_freq] s[ids] = obj return s
def _normalize_seqs(s, t)
Normalize to RPM
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# use pybedtools to keep valid positions # intersect option with -b bigger_cluster_loci a = pybedtools.BedTool(bam_in) b = pybedtools.BedTool(precursors) c = a.intersect(b, u=True) out_file = utils.splitext_plus(op.basename(bam_in))[0] + "_clean.bam" c.saveas(out_file) return op.abs...
def prepare_bam(bam_in, precursors)
Clean BAM file to keep only position inside the bigger cluster
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new_bed = utils.splitext_plus(bed_file)[0] + '_order.bed' with open(bed_file) as in_handle: with open(new_bed, 'w') as out_handle: for line in in_handle: cols = line.strip().split("\t") cols[3] = _select_anno(cols[3]) + "_" + cols[4] cols[...
def _reorder_columns(bed_file)
Reorder columns to be compatible with CoRaL
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new_cov = utils.splitext_plus(cov_file)[0] + '_fix.cov' with open(cov_file) as in_handle: with open(new_cov, 'w') as out_handle: for line in in_handle: cols = line.strip().split("\t") cols[4] = cols[6] print("\t".join(cols[0:6]), file=out_...
def _fix_score_column(cov_file)
Move counts to score columns in bed file
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bed_file = _reorder_columns(bed_file) counts_reads_cmd = ("coverageBed -s -counts -b {bam_in} " "-a {bed_file} | sort -k4,4 " "> {out_dir}/loci.cov") # with tx_tmpdir() as temp_dir: with utils.chdir(out_dir): run(counts_reads_cmd.format(min_tr...
def detect_regions(bam_in, bed_file, out_dir, prefix)
Detect regions using first CoRaL module
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0.987955
new_cov = op.join(op.dirname(cov_file), 'feat_antisense.txt') with open(cov_file) as in_handle: with open(new_cov, 'w') as out_handle: print("name\tantisense", file=out_handle, end="") for line in in_handle: cols = line.strip().split("\t") col...
def _order_antisense_column(cov_file, min_reads)
Move counts to score columns in bed file
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1.02495
data = Counter() a = pybedtools.BedTool(bam_in) b = pybedtools.BedTool(loci_file) c = a.intersect(b, s=True, bed=True, wo=True) for line in c: end = int(line[1]) + 1 + int(line[2]) if line[5] == "+" else int(line[1]) + 1 start = int(line[1]) + 1 if line[5] == "+" else int(line[1...
def _reads_per_position(bam_in, loci_file, out_dir)
Create input for compute entropy
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