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1 #!/usr/bin/env python
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2
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3 import optparse, os, shutil, subprocess, sys, tempfile
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4
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5 def stop_err( msg ):
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6 sys.stderr.write( '%s\n' % msg )
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7 sys.exit()
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8
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9 def __main__():
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10 #Parse Command Line
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11 parser = optparse.OptionParser()
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12 parser.add_option( '-g', dest='ref_annotation', help='An optional "reference" annotation GTF. Each sample is matched against this file, and sample isoforms are tagged as overlapping, matching, or novel where appropriate. See the refmap and tmap output file descriptions below.' )
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13 parser.add_option( '-s', dest='use_seq_data', action="store_true", help='Causes cuffmerge to look into for fasta files with the underlying genomic sequences (one file per contig) against which your reads were aligned for some optional classification functions. For example, Cufflinks transcripts consisting mostly of lower-case bases are classified as repeats. Note that <seq_dir> must contain one fasta file per reference chromosome, and each file must be named after the chromosome, and have a .fa or .fasta extension.')
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14 parser.add_option( '-p', '--num-threads', dest='num_threads', help='Use this many threads to align reads. The default is 1.' )
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15
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16 # Wrapper / Galaxy options.
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17 parser.add_option( '', '--index', dest='index', help='The path of the reference genome' )
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18 parser.add_option( '', '--ref_file', dest='ref_file', help='The reference dataset from the history' )
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19
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20 # Outputs.
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21 parser.add_option( '', '--merged-transcripts', dest='merged_transcripts' )
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22 parser.add_option( '--min-isoform-fraction', dest='min_isoform_fraction' )
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23
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24 (options, args) = parser.parse_args()
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25
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26 # output version # of tool
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27 try:
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28 tmp = tempfile.NamedTemporaryFile().name
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29 tmp_stdout = open( tmp, 'wb' )
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30 proc = subprocess.Popen( args='cuffmerge -v 2>&1', shell=True, stdout=tmp_stdout )
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31 tmp_stdout.close()
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32 returncode = proc.wait()
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33 stdout = None
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34 for line in open( tmp_stdout.name, 'rb' ):
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35 if line.lower().find( 'merge_cuff_asms v' ) >= 0:
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36 stdout = line.strip()
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37 break
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38 if stdout:
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39 sys.stdout.write( '%s\n' % stdout )
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40 else:
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41 raise Exception
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42 except:
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43 sys.stdout.write( 'Could not determine Cuffmerge version\n' )
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44
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45 # Set/link to sequence file.
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46 if options.use_seq_data:
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47 if options.ref_file:
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48 # Sequence data from history.
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49 # Create symbolic link to ref_file so that index will be created in working directory.
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50 seq_path = "ref.fa"
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51 os.symlink( options.ref_file, seq_path )
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52 else:
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53 if not os.path.exists( options.index ):
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54 stop_err( 'Reference genome %s not present, request it by reporting this error.' % options.index )
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55 seq_path = options.index
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56
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57 # Build command.
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58
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59 # Base.
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60 cmd = "cuffmerge -o cm_output "
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61
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62 # Add options.
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63 if options.num_threads:
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64 cmd += ( " -p %i " % int ( options.num_threads ) )
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65 if options.ref_annotation:
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66 cmd += " -g %s " % options.ref_annotation
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67 if options.use_seq_data:
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68 cmd += " -s %s " % seq_path
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69 if options.min_isoform_fraction:
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70 cmd += " --min-isoform-fraction %s " % (options.min_isoform_fraction)
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71 # Add input files to a file.
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72 inputs_file_name = tempfile.NamedTemporaryFile( dir="." ).name
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73 inputs_file = open( inputs_file_name, 'w' )
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74 for arg in args:
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75 inputs_file.write( arg + "\n" )
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76 inputs_file.close()
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77 cmd += inputs_file_name
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78
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79 # Debugging.
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80 print cmd
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81
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82 # Run command.
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83 try:
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84 tmp_name = tempfile.NamedTemporaryFile( dir="." ).name
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85 tmp_stderr = open( tmp_name, 'wb' )
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86 proc = subprocess.Popen( args=cmd, shell=True, stderr=tmp_stderr.fileno() )
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87 returncode = proc.wait()
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88 tmp_stderr.close()
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89
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90 # Get stderr, allowing for case where it's very large.
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91 tmp_stderr = open( tmp_name, 'rb' )
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92 stderr = ''
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93 buffsize = 1048576
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94 try:
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95 while True:
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96 stderr += tmp_stderr.read( buffsize )
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97 if not stderr or len( stderr ) % buffsize != 0:
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98 break
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99 except OverflowError:
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100 pass
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101 tmp_stderr.close()
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102
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103 # Error checking.
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104 if returncode != 0:
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105 raise Exception, stderr
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106
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107 if len( open( "cm_output/merged.gtf", 'rb' ).read().strip() ) == 0:
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108 raise Exception, 'The output file is empty, there may be an error with your input file or settings.'
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109
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110 # Copy outputs.
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111 shutil.copyfile( "cm_output/merged.gtf" , options.merged_transcripts )
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112
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113 except Exception, e:
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114 stop_err( 'Error running cuffmerge. ' + str( e ) )
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115
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116 if __name__=="__main__": __main__()
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