Mercurial > repos > rnateam > bctools
view merge_pcr_duplicates.py @ 50:0b9aab6aaebf draft
Uploaded 16cfcafe8b42055c5dd64e62c42b82b455027a40
author | rnateam |
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date | Tue, 26 Jan 2016 04:38:27 -0500 |
parents | 570a7de9f151 |
children | 4bedd35bcdff |
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#!/usr/bin/env python import argparse import logging from sys import stdout import pandas as pd from subprocess import check_call from shutil import rmtree from tempfile import mkdtemp from os.path import isfile # avoid ugly python IOError when stdout output is piped into another program # and then truncated (such as piping to head) from signal import signal, SIGPIPE, SIG_DFL signal(SIGPIPE, SIG_DFL) tool_description = """ Merge PCR duplicates according to random barcode library. Barcodes containing uncalled base 'N' are removed. By default output is written to stdout. Input: * bed6 file containing alignments with fastq read-id in name field * fastq library of random barcodes Output: * bed6 file with random barcode in name field and number of PCR duplicates as score, sorted by fields chrom, start, stop, strand, name Example usage: - read PCR duplicates from file duplicates.bed and write merged results to file merged.bed: merge_pcr_duplicates.py duplicates.bed bclibrary.fa --out merged.bed """ epilog = """ Author: Daniel Maticzka Copyright: 2015 License: Apache Email: maticzkd@informatik.uni-freiburg.de Status: Testing """ # parse command line arguments parser = argparse.ArgumentParser(description=tool_description, epilog=epilog, formatter_class=argparse.RawDescriptionHelpFormatter) # positional arguments parser.add_argument( "alignments", help="Path to bed6 file containing alignments.") parser.add_argument( "bclib", help="Path to fastq barcode library.") # optional arguments parser.add_argument( "-o", "--outfile", help="Write results to this file.") # misc arguments parser.add_argument( "-v", "--verbose", help="Be verbose.", action="store_true") parser.add_argument( "-d", "--debug", help="Print lots of debugging information", action="store_true") parser.add_argument( '--version', action='version', version='0.2.0') args = parser.parse_args() if args.debug: logging.basicConfig(level=logging.DEBUG, format="%(asctime)s - %(filename)s - %(levelname)s - %(message)s") elif args.verbose: logging.basicConfig(level=logging.INFO, format="%(filename)s - %(levelname)s - %(message)s") else: logging.basicConfig(format="%(filename)s - %(levelname)s - %(message)s") logging.info("Parsed arguments:") logging.info(" alignments: '{}'".format(args.alignments)) logging.info(" bclib: '{}'".format(args.bclib)) if args.outfile: logging.info(" outfile: enabled writing to file") logging.info(" outfile: '{}'".format(args.outfile)) logging.info("") # see if alignments are empty and the tool can quit n_alns = sum(1 for line in open(args.alignments)) if n_alns == 0: logging.warning("WARNING: Working on empty set of alignments, writing empty output.") eventalnout = (open(args.outfile, "w") if args.outfile is not None else stdout) eventalnout.close() exit(0) # check input filenames if not isfile(args.bclib): raise Exception("ERROR: barcode library '{}' not found.") if not isfile(args.alignments): raise Exception("ERROR: alignments '{}' not found.") try: tmpdir = mkdtemp() logging.debug("tmpdir: " + tmpdir) # prepare barcode library syscall1 = "cat " + args.bclib + " | awk 'BEGIN{OFS=\"\\t\"}NR%4==1{gsub(/^@/,\"\"); id=$1}NR%4==2{bc=$1}NR%4==3{print id,bc}' | sort -k1,1 > " + tmpdir + "/bclib.csv" check_call(syscall1, shell=True) # prepare alinments syscall2 = "cat " + args.alignments + " | awk -F \"\\t\" 'BEGIN{OFS=\"\\t\"}{split($4, a, \" \"); $4 = a[1]; print}'| sort -k4,4 > " + tmpdir + "/alns.csv" check_call(syscall2, shell=True) # join barcode library and alignments syscall3 = "join -1 1 -2 4 " + tmpdir + "/bclib.csv " + tmpdir + "/alns.csv " + " | awk 'BEGIN{OFS=\"\\t\"}{print $3,$4,$5,$2,$6,$7}' > " + tmpdir + "/bcalib.csv" check_call(syscall3, shell=True) # get alignments combined with barcodes bcalib = pd.read_csv( tmpdir + "/bcalib.csv", sep="\t", names=["chrom", "start", "stop", "bc", "score", "strand"]) finally: logging.debug("removed tmpdir: " + tmpdir) rmtree(tmpdir) # fail if alignments given but combined library is empty if bcalib.empty: raise Exception("ERROR: no common entries for alignments and barcode library found. Please check your input files.") # warn if not all alignments could be assigned a barcode n_bcalib = len(bcalib.index) if n_bcalib < n_alns: logging.warning( "{} of {} alignments could not be associated with a random barcode.".format(n_alns - n_bcalib, n_alns)) # remove entries with barcodes that has uncalled base N bcalib_cleaned = bcalib.drop(bcalib[bcalib.bc.str.contains("N")].index) n_bcalib_cleaned = len(bcalib_cleaned) # if n_bcalib_cleaned < n_bcalib: # msg = "{} of {} alignments had random barcodes containing uncalled bases and were dropped.".format( # n_bcalib - n_bcalib_cleaned, n_bcalib) # if n_bcalib_cleaned < (0.8 * n_bcalib): # logging.warning(msg) # else: # logging.info(msg) # count and merge pcr duplicates # grouping sorts by keys, so the ouput will be properly sorted merged = bcalib_cleaned.groupby(['chrom', 'start', 'stop', 'strand', 'bc']).size().reset_index() merged.rename(columns={0: 'ndupes'}, copy=False, inplace=True) # write coordinates of crosslinking event alignments eventalnout = (open(args.outfile, "w") if args.outfile is not None else stdout) merged.to_csv( eventalnout, columns=['chrom', 'start', 'stop', 'bc', 'ndupes', 'strand'], sep="\t", index=False, header=False) eventalnout.close()