annotate bowtie_genome_wrapper/bowtie_genomic_wrapper.py @ 2:7e3bd576aa7c draft default tip

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author triasteran
date Tue, 26 Apr 2022 12:21:05 +0000
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1 #!/usr/bin/env python
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2
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3 """
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4 Runs Bowtie on single-end or paired-end data.
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5
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6 usage: bowtie_wrapper.py [options]
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7 -t, --threads=t: The number of threads to run
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8 -o, --output=o: The output file
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9 --output_unmapped_reads=: File name for unmapped reads (single-end)
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10 --output_unmapped_reads_l=: File name for unmapped reads (left, paired-end)
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11 --output_unmapped_reads_r=: File name for unmapped reads (right, paired-end)
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12 --output_suppressed_reads=: File name for suppressed reads because of max setting (single-end)
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13 --output_suppressed_reads_l=: File name for suppressed reads because of max setting (left, paired-end)
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14 --output_suppressed_reads_r=: File name for suppressed reads because of max setting (right, paired-end)
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15 --output_mapping_stats=: File name for mapping statistics (output on stderr by bowtie)
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16 -i, --input1=i: The (forward or single-end) reads file in Sanger FASTQ format
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17 -I, --input2=I: The reverse reads file in Sanger FASTQ format
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18 -4, --dataType=4: The type of data (SOLiD or Solexa)
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19 -2, --paired=2: Whether the data is single- or paired-end
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20 -g, --genomeSource=g: The type of reference provided
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21 -r, --ref=r: The reference genome to use or index
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22 -s, --skip=s: Skip the first n reads
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23 -a, --alignLimit=a: Only align the first n reads
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24 -T, --trimH=T: Trim n bases from high-quality (left) end of each read before alignment
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25 -L, --trimL=L: Trim n bases from low-quality (right) end of each read before alignment
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26 -m, --mismatchSeed=m: Maximum number of mismatches permitted in the seed
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27 -M, --mismatchQual=M: Maximum permitted total of quality values at mismatched read positions
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28 -l, --seedLen=l: Seed length
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29 -n, --rounding=n: Whether or not to round to the nearest 10 and saturating at 30
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30 -P, --maxMismatches=P: Maximum number of mismatches for -v alignment mode
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31 -w, --tryHard=: Whether or not to try as hard as possible to find valid alignments when they exist
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32 -V, --allValAligns=V: Whether or not to report all valid alignments per read or pair
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33 -v, --valAlign=v: Report up to n valid alignments per read or pair
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34 -G, --suppressAlign=G: Suppress all alignments for a read if more than n reportable alignments exist
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35 -b, --best=b: Whether or not to make Bowtie guarantee that reported singleton alignments are 'best' in terms of stratum and in terms of the quality values at the mismatched positions
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36 -B, --maxBacktracks=B: Maximum number of backtracks permitted when aligning a read
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37 -R, --strata=R: Whether or not to report only those alignments that fall in the best stratum if many valid alignments exist and are reportable
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38 -j, --minInsert=j: Minimum insert size for valid paired-end alignments
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39 -J, --maxInsert=J: Maximum insert size for valid paired-end alignments
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40 -O, --mateOrient=O: The upstream/downstream mate orientation for valid paired-end alignment against the forward reference strand
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41 -A, --maxAlignAttempt=A: Maximum number of attempts Bowtie will make to match an alignment for one mate with an alignment for the opposite mate
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42 -f, --forwardAlign=f: Whether or not to attempt to align the forward reference strand
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43 -E, --reverseAlign=E: Whether or not to attempt to align the reverse-complement reference strand
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44 -F, --offrate=F: Override the offrate of the index to n
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45 -8, --snpphred=8: SNP penalty on Phred scale
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46 -6, --snpfrac=6: Fraction of sites expected to be SNP sites
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47 -7, --keepends=7: Keep extreme-end nucleotides and qualities
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48 -S, --seed=S: Seed for pseudo-random number generator
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49 -C, --params=C: Whether to use default or specified parameters
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50 -u, --iautoB=u: Automatic or specified behavior
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51 -K, --ipacked=K: Whether or not to use a packed representation for DNA strings
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52 -Q, --ibmax=Q: Maximum number of suffixes allowed in a block
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53 -Y, --ibmaxdivn=Y: Maximum number of suffixes allowed in a block as a fraction of the length of the reference
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54 -D, --idcv=D: The period for the difference-cover sample
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55 -U, --inodc=U: Whether or not to disable the use of the difference-cover sample
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56 -y, --inoref=y: Whether or not to build the part of the reference index used only in paired-end alignment
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57 -z, --ioffrate=z: How many rows get marked during annotation of some or all of the Burrows-Wheeler rows
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58 -W, --iftab=W: The size of the lookup table used to calculate an initial Burrows-Wheeler range with respect to the first n characters of the query
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59 -X, --intoa=X: Whether or not to convert Ns in the reference sequence to As
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60 -N, --iendian=N: Endianness to use when serializing integers to the index file
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61 -Z, --iseed=Z: Seed for the pseudorandom number generator
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62 -x, --indexSettings=x: Whether or not indexing options are to be set
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63 -H, --suppressHeader=H: Suppress header
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64 --do_not_build_index: Flag to specify that provided file is already indexed and to just use 'as is'
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65 """
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66
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67 import optparse
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68 import os
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69 import shutil
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70 import subprocess
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71 import sys
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72 import tempfile
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73
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74 # Allow more than Sanger encoded variants
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75 DEFAULT_ASCII_ENCODING = '--phred33-quals'
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76 GALAXY_FORMAT_TO_QUALITY_SCORE_ENCODING_ARG = {'fastqsanger': '--phred33-quals', 'fastqillumina': '--phred64-quals', 'fastqsolexa': '--solexa-quals'}
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77 # FIXME: Integer quality scores are supported only when the '--integer-quals' argument is specified to bowtie; this is not currently able to be set in the tool/wrapper/config
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78
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79
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80 def stop_err( msg ):
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81 sys.exit('%s\n' % msg)
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82
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83
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84 def __main__():
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85 parser = optparse.OptionParser()
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86 parser.add_option( '-t', '--threads', dest='threads', help='The number of threads to run' )
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87 parser.add_option( '-o', '--output', dest='output', help='The output file' )
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88 parser.add_option( '', '--output_unmapped_reads', dest='output_unmapped_reads', help='File name for unmapped reads (single-end)' )
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89 parser.add_option( '', '--output_unmapped_reads_l', dest='output_unmapped_reads_l', help='File name for unmapped reads (left, paired-end)' )
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90 parser.add_option( '', '--output_unmapped_reads_r', dest='output_unmapped_reads_r', help='File name for unmapped reads (right, paired-end)' )
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91 parser.add_option( '', '--output_suppressed_reads', dest='output_suppressed_reads', help='File name for suppressed reads because of max setting (single-end)' )
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92 parser.add_option( '', '--output_suppressed_reads_l', dest='output_suppressed_reads_l', help='File name for suppressed reads because of max setting (left, paired-end)' )
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93 parser.add_option( '', '--output_suppressed_reads_r', dest='output_suppressed_reads_r', help='File name for suppressed reads because of max setting (right, paired-end)' )
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94 parser.add_option( '', '--output_mapping_stats', dest='output_mapping_stats', help='File for mapping statistics (i.e. stderr from bowtie)' )
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95 parser.add_option( '-4', '--dataType', dest='dataType', help='The type of data (SOLiD or Solexa)' )
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96 parser.add_option( '-i', '--input1', dest='input1', help='The (forward or single-end) reads file in Sanger FASTQ format' )
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97 parser.add_option( '-I', '--input2', dest='input2', help='The reverse reads file in Sanger FASTQ format' )
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98 parser.add_option( '-2', '--paired', dest='paired', help='Whether the data is single- or paired-end' )
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99 parser.add_option( '-g', '--genomeSource', dest='genomeSource', help='The type of reference provided' )
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100 parser.add_option( '-r', '--ref', dest='ref', help='The reference genome to use or index' )
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101 parser.add_option( '-s', '--skip', dest='skip', help='Skip the first n reads' )
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102 parser.add_option( '-a', '--alignLimit', dest='alignLimit', help='Only align the first n reads' )
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103 parser.add_option( '-T', '--trimH', dest='trimH', help='Trim n bases from high-quality (left) end of each read before alignment' )
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104 parser.add_option( '-L', '--trimL', dest='trimL', help='Trim n bases from low-quality (right) end of each read before alignment' )
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105 parser.add_option( '-m', '--mismatchSeed', dest='mismatchSeed', help='Maximum number of mismatches permitted in the seed' )
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106 parser.add_option( '-M', '--mismatchQual', dest='mismatchQual', help='Maximum permitted total of quality values at mismatched read positions' )
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107 parser.add_option( '-l', '--seedLen', dest='seedLen', help='Seed length' )
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108 parser.add_option( '-n', '--rounding', dest='rounding', help='Whether or not to round to the nearest 10 and saturating at 30' )
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109 parser.add_option( '-P', '--maxMismatches', dest='maxMismatches', help='Maximum number of mismatches for -v alignment mode' )
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110 parser.add_option( '-w', '--tryHard', dest='tryHard', help='Whether or not to try as hard as possible to find valid alignments when they exist' )
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111 parser.add_option( '-V', '--allValAligns', dest='allValAligns', help='Whether or not to report all valid alignments per read or pair' )
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112 parser.add_option( '-v', '--valAlign', dest='valAlign', help='Report up to n valid alignments per read or pair' )
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113 parser.add_option( '-G', '--suppressAlign', dest='suppressAlign', help='Suppress all alignments for a read if more than n reportable alignments exist' )
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114 parser.add_option( '-b', '--best', dest='best', help="Whether or not to make Bowtie guarantee that reported singleton alignments are 'best' in terms of stratum and in terms of the quality values at the mismatched positions" )
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115 parser.add_option( '-B', '--maxBacktracks', dest='maxBacktracks', help='Maximum number of backtracks permitted when aligning a read' )
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116 parser.add_option( '-R', '--strata', dest='strata', help='Whether or not to report only those alignments that fall in the best stratum if many valid alignments exist and are reportable' )
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117 parser.add_option( '-j', '--minInsert', dest='minInsert', help='Minimum insert size for valid paired-end alignments' )
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118 parser.add_option( '-J', '--maxInsert', dest='maxInsert', help='Maximum insert size for valid paired-end alignments' )
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119 parser.add_option( '-O', '--mateOrient', dest='mateOrient', help='The upstream/downstream mate orientation for valid paired-end alignment against the forward reference strand' )
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120 parser.add_option( '-A', '--maxAlignAttempt', dest='maxAlignAttempt', help='Maximum number of attempts Bowtie will make to match an alignment for one mate with an alignment for the opposite mate' )
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121 parser.add_option( '-f', '--forwardAlign', dest='forwardAlign', help='Whether or not to attempt to align the forward reference strand' )
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122 parser.add_option( '-E', '--reverseAlign', dest='reverseAlign', help='Whether or not to attempt to align the reverse-complement reference strand' )
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123 parser.add_option( '-F', '--offrate', dest='offrate', help='Override the offrate of the index to n' )
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124 parser.add_option( '-S', '--seed', dest='seed', help='Seed for pseudo-random number generator' )
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125 parser.add_option( '-8', '--snpphred', dest='snpphred', help='SNP penalty on Phred scale' )
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126 parser.add_option( '-6', '--snpfrac', dest='snpfrac', help='Fraction of sites expected to be SNP sites' )
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127 parser.add_option( '-7', '--keepends', dest='keepends', help='Keep extreme-end nucleotides and qualities' )
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128 parser.add_option( '-C', '--params', dest='params', help='Whether to use default or specified parameters' )
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129 parser.add_option( '-u', '--iautoB', dest='iautoB', help='Automatic or specified behavior' )
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130 parser.add_option( '-K', '--ipacked', dest='ipacked', help='Whether or not to use a packed representation for DNA strings' )
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131 parser.add_option( '-Q', '--ibmax', dest='ibmax', help='Maximum number of suffixes allowed in a block' )
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132 parser.add_option( '-Y', '--ibmaxdivn', dest='ibmaxdivn', help='Maximum number of suffixes allowed in a block as a fraction of the length of the reference' )
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133 parser.add_option( '-D', '--idcv', dest='idcv', help='The period for the difference-cover sample' )
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134 parser.add_option( '-U', '--inodc', dest='inodc', help='Whether or not to disable the use of the difference-cover sample' )
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135 parser.add_option( '-y', '--inoref', dest='inoref', help='Whether or not to build the part of the reference index used only in paired-end alignment' )
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136 parser.add_option( '-z', '--ioffrate', dest='ioffrate', help='How many rows get marked during annotation of some or all of the Burrows-Wheeler rows' )
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137 parser.add_option( '-W', '--iftab', dest='iftab', help='The size of the lookup table used to calculate an initial Burrows-Wheeler range with respect to the first n characters of the query' )
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138 parser.add_option( '-X', '--intoa', dest='intoa', help='Whether or not to convert Ns in the reference sequence to As' )
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139 parser.add_option( '-N', '--iendian', dest='iendian', help='Endianness to use when serializing integers to the index file' )
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140 parser.add_option( '-Z', '--iseed', dest='iseed', help='Seed for the pseudorandom number generator' )
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141 parser.add_option( '-x', '--indexSettings', dest='index_settings', help='Whether or not indexing options are to be set' )
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142 parser.add_option( '-H', '--suppressHeader', dest='suppressHeader', help='Suppress header' )
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143 parser.add_option( '--galaxy_input_format', dest='galaxy_input_format', default="fastqsanger", help='galaxy input format' )
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144 parser.add_option( '--do_not_build_index', dest='do_not_build_index', action="store_true", default=False, help='Flag to specify that provided file is already indexed, use as is' )
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145 (options, args) = parser.parse_args()
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146 if options.mismatchSeed and options.maxMismatches:
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147 parser.error("options --mismatchSeed and --maxMismatches are mutually exclusive")
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
148 stdout = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
149
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
150 # make temp directory for placement of indices and copy reference file there if necessary
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
151 tmp_index_dir = tempfile.mkdtemp()
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
152 # get type of data (solid or solexa)
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
153 if options.dataType == 'solid':
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triasteran
parents:
diff changeset
154 colorspace = '-C'
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triasteran
parents:
diff changeset
155 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
156 colorspace = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
157 # index if necessary
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
158 if options.genomeSource == 'history' and not options.do_not_build_index:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
159 # set up commands
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
160 if options.index_settings == 'indexPreSet':
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
161 indexing_cmds = '%s' % colorspace
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
162 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
163 try:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
164 if options.iautoB and options.iautoB == 'set':
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
165 iautoB = '--noauto'
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
166 else:
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triasteran
parents:
diff changeset
167 iautoB = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
168 if options.ipacked and options.ipacked == 'packed':
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triasteran
parents:
diff changeset
169 ipacked = '--packed'
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
170 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
171 ipacked = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
172 if options.ibmax and int( options.ibmax ) >= 1:
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triasteran
parents:
diff changeset
173 ibmax = '--bmax %s' % options.ibmax
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
174 else:
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triasteran
parents:
diff changeset
175 ibmax = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
176 if options.ibmaxdivn and int( options.ibmaxdivn ) >= 0:
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triasteran
parents:
diff changeset
177 ibmaxdivn = '--bmaxdivn %s' % options.ibmaxdivn
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
178 else:
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triasteran
parents:
diff changeset
179 ibmaxdivn = ''
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triasteran
parents:
diff changeset
180 if options.idcv and int( options.idcv ) >= 3:
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triasteran
parents:
diff changeset
181 idcv = '--dcv %s' % options.idcv
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
182 else:
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triasteran
parents:
diff changeset
183 idcv = ''
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triasteran
parents:
diff changeset
184 if options.inodc and options.inodc == 'nodc':
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triasteran
parents:
diff changeset
185 inodc = '--nodc'
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triasteran
parents:
diff changeset
186 else:
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triasteran
parents:
diff changeset
187 inodc = ''
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triasteran
parents:
diff changeset
188 if options.inoref and options.inoref == 'noref':
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triasteran
parents:
diff changeset
189 inoref = '--noref'
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
190 else:
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triasteran
parents:
diff changeset
191 inoref = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
192 if options.iftab and int( options.iftab ) >= 1:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
193 iftab = '--ftabchars %s' % options.iftab
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
194 else:
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triasteran
parents:
diff changeset
195 iftab = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
196 if options.intoa and options.intoa == 'yes':
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triasteran
parents:
diff changeset
197 intoa = '--ntoa'
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
198 else:
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triasteran
parents:
diff changeset
199 intoa = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
200 if options.iendian and options.iendian == 'big':
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
201 iendian = '--big'
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
202 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
203 iendian = '--little'
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
204 if options.iseed and int( options.iseed ) > 0:
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triasteran
parents:
diff changeset
205 iseed = '--seed %s' % options.iseed
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
206 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
207 iseed = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
208 indexing_cmds = '%s %s %s %s %s %s %s --offrate %s %s %s %s %s %s' % \
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
209 ( iautoB, ipacked, ibmax, ibmaxdivn, idcv, inodc,
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
210 inoref, options.ioffrate, iftab, intoa, iendian,
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
211 iseed, colorspace )
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
212 except ValueError as e:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
213 # clean up temp dir
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
214 if os.path.exists( tmp_index_dir ):
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
215 shutil.rmtree( tmp_index_dir )
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
216 stop_err( "Something is wrong with the indexing parameters and the indexing and alignment could not be run. Make sure you don't have any non-numeric values where they should be numeric.\n" + str( e ) )
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
217 ref_file = tempfile.NamedTemporaryFile( dir=tmp_index_dir )
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
218 ref_file_name = ref_file.name
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
219 ref_file.close()
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
220 os.symlink( options.ref, ref_file_name )
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
221 cmd1 = 'bowtie-build %s -f %s %s' % ( indexing_cmds, ref_file_name, ref_file_name )
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
222 try:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
223 tmp = tempfile.NamedTemporaryFile( dir=tmp_index_dir ).name
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
224 with open(tmp, 'w') as tmp_stderr:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
225 returncode = subprocess.call(args=cmd1, shell=True, cwd=tmp_index_dir, stderr=tmp_stderr.fileno())
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
226 if returncode != 0:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
227 # get stderr, allowing for case where it's very large
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
228 stderr = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
229 buffsize = 1048576
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
230 with open(tmp, 'r') as tmp_stderr:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
231 try:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
232 while True:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
233 stderr += tmp_stderr.read(buffsize)
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
234 if not stderr or len(stderr) % buffsize != 0:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
235 break
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
236 except OverflowError:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
237 pass
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
238 raise Exception(stderr)
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
239 except Exception as e:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
240 # clean up temp dir
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
241 if os.path.exists( tmp_index_dir ):
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
242 shutil.rmtree( tmp_index_dir )
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
243 stop_err( 'Error indexing reference sequence\n' + str( e ) )
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
244 stdout += 'File indexed. '
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
245 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
246 ref_file_name = options.ref
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
247 # set up aligning and generate aligning command options
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
248 # automatically set threads in both cases
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
249 tmp_suppressed_file_name = None
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
250 tmp_unmapped_file_name = None
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
251 if options.suppressHeader == 'true':
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
252 suppressHeader = '--sam-nohead'
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
253 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
254 suppressHeader = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
255 if options.maxInsert and int( options.maxInsert ) > 0:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
256 maxInsert = '-X %s' % options.maxInsert
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
257 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
258 maxInsert = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
259 if options.mateOrient:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
260 mateOrient = '--%s' % options.mateOrient
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
261 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
262 mateOrient = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
263 quality_score_encoding = GALAXY_FORMAT_TO_QUALITY_SCORE_ENCODING_ARG.get( options.galaxy_input_format, DEFAULT_ASCII_ENCODING )
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
264 if options.params == 'preSet':
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
265 aligning_cmds = '-q %s %s -p %s -S %s %s %s ' % \
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
266 ( maxInsert, mateOrient, options.threads, suppressHeader, colorspace, quality_score_encoding )
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
267 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
268 try:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
269 if options.skip and int( options.skip ) > 0:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
270 skip = '-s %s' % options.skip
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
271 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
272 skip = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
273 if options.alignLimit and int( options.alignLimit ) >= 0:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
274 alignLimit = '-u %s' % options.alignLimit
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
275 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
276 alignLimit = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
277 if options.trimH and int( options.trimH ) > 0:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
278 trimH = '-5 %s' % options.trimH
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
279 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
280 trimH = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
281 if options.trimL and int( options.trimL ) > 0:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
282 trimL = '-3 %s' % options.trimL
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
283 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
284 trimL = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
285 if options.maxMismatches and (options.maxMismatches == '0' or options.maxMismatches == '1' or
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
286 options.maxMismatches == '2' or options.maxMismatches == '3'):
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
287 maxMismatches = '-v %s' % options.maxMismatches
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
288 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
289 maxMismatches = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
290 if options.mismatchSeed and (options.mismatchSeed == '0' or options.mismatchSeed == '1' or
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
291 options.mismatchSeed == '2' or options.mismatchSeed == '3'):
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
292 mismatchSeed = '-n %s' % options.mismatchSeed
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
293 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
294 mismatchSeed = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
295 if options.mismatchQual and int( options.mismatchQual ) >= 1:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
296 mismatchQual = '-e %s' % options.mismatchQual
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
297 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
298 mismatchQual = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
299 if options.seedLen and int( options.seedLen ) >= 5:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
300 seedLen = '-l %s' % options.seedLen
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
301 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
302 seedLen = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
303 if options.rounding == 'noRound':
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
304 rounding = '--nomaqround'
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
305 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
306 rounding = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
307 if options.minInsert and int( options.minInsert ) > 0:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
308 minInsert = '-I %s' % options.minInsert
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
309 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
310 minInsert = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
311 if options.maxAlignAttempt and int( options.maxAlignAttempt ) >= 0:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
312 maxAlignAttempt = '--pairtries %s' % options.maxAlignAttempt
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
313 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
314 maxAlignAttempt = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
315 if options.forwardAlign == 'noForward':
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
316 forwardAlign = '--nofw'
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
317 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
318 forwardAlign = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
319 if options.reverseAlign == 'noReverse':
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
320 reverseAlign = '--norc'
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
321 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
322 reverseAlign = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
323 if options.maxBacktracks and int( options.maxBacktracks ) > 0 and \
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
324 ( options.mismatchSeed == '2' or options.mismatchSeed == '3' ):
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
325 maxBacktracks = '--maxbts %s' % options.maxBacktracks
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
326 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
327 maxBacktracks = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
328 if options.tryHard == 'doTryHard':
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
329 tryHard = '-y'
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
330 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
331 tryHard = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
332 if options.valAlign and int( options.valAlign ) >= 0:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
333 valAlign = '-k %s' % options.valAlign
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
334 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
335 valAlign = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
336 if options.allValAligns == 'doAllValAligns':
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
337 allValAligns = '-a'
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
338 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
339 allValAligns = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
340 if options.suppressAlign and int( options.suppressAlign ) >= 0:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
341 suppressAlign = '-m %s' % options.suppressAlign
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
342 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
343 suppressAlign = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
344 if options.best == 'doBest':
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
345 best = '--best'
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
346 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
347 best = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
348 if options.strata == 'doStrata':
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
349 strata = '--strata'
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
350 else:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
351 strata = ''
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
352 if options.offrate and int( options.offrate ) >= 0:
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parents:
diff changeset
353 offrate = '-o %s' % options.offrate
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parents:
diff changeset
354 else:
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parents:
diff changeset
355 offrate = ''
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parents:
diff changeset
356 if options.seed and int( options.seed ) >= 0:
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parents:
diff changeset
357 seed = '--seed %s' % options.seed
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parents:
diff changeset
358 else:
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parents:
diff changeset
359 seed = ''
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parents:
diff changeset
360 if options.paired == 'paired':
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parents:
diff changeset
361 if options.output_unmapped_reads_l and options.output_unmapped_reads_r:
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parents:
diff changeset
362 tmp_unmapped_file = tempfile.NamedTemporaryFile( dir=tmp_index_dir, suffix='.fastq' )
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parents:
diff changeset
363 tmp_unmapped_file_name = tmp_unmapped_file.name
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parents:
diff changeset
364 tmp_unmapped_file.close()
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parents:
diff changeset
365 output_unmapped_reads = '--un %s' % tmp_unmapped_file_name
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parents:
diff changeset
366 else:
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parents:
diff changeset
367 output_unmapped_reads = ''
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parents:
diff changeset
368 if options.output_suppressed_reads:
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parents:
diff changeset
369 tmp_suppressed_file = tempfile.NamedTemporaryFile( dir=tmp_index_dir, suffix='.fastq' )
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parents:
diff changeset
370 tmp_suppressed_file_name = tmp_suppressed_file.name
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parents:
diff changeset
371 tmp_suppressed_file.close()
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parents:
diff changeset
372 output_suppressed_reads = '--max %s' % tmp_suppressed_file_name
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parents:
diff changeset
373 else:
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parents:
diff changeset
374 output_suppressed_reads = ''
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parents:
diff changeset
375 else:
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triasteran
parents:
diff changeset
376 if options.output_unmapped_reads:
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parents:
diff changeset
377 output_unmapped_reads = '--un %s' % options.output_unmapped_reads
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parents:
diff changeset
378 else:
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parents:
diff changeset
379 output_unmapped_reads = ''
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triasteran
parents:
diff changeset
380 if options.output_suppressed_reads:
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parents:
diff changeset
381 output_suppressed_reads = '--max %s' % options.output_suppressed_reads
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parents:
diff changeset
382 else:
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parents:
diff changeset
383 output_suppressed_reads = ''
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triasteran
parents:
diff changeset
384 snpfrac = ''
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parents:
diff changeset
385 if options.snpphred and int( options.snpphred ) >= 0:
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parents:
diff changeset
386 snpphred = '--snpphred %s' % options.snpphred
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parents:
diff changeset
387 else:
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parents:
diff changeset
388 snpphred = ''
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parents:
diff changeset
389 if options.snpfrac and float( options.snpfrac ) >= 0:
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parents:
diff changeset
390 snpfrac = '--snpfrac %s' % options.snpfrac
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triasteran
parents:
diff changeset
391 if options.keepends and options.keepends == 'doKeepends':
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parents:
diff changeset
392 keepends = '--col-keepends'
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parents:
diff changeset
393 else:
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parents:
diff changeset
394 keepends = ''
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parents:
diff changeset
395 aligning_cmds = '-q %s %s -p %s -S %s %s %s %s %s %s %s %s %s %s %s %s ' \
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parents:
diff changeset
396 '%s %s %s %s %s %s %s %s %s %s %s %s %s %s %s %s %s %s ' % \
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triasteran
parents:
diff changeset
397 ( maxInsert, mateOrient, options.threads, suppressHeader,
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
398 colorspace, skip, alignLimit, trimH, trimL, maxMismatches,
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
399 mismatchSeed, mismatchQual, seedLen, rounding, minInsert,
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
400 maxAlignAttempt, forwardAlign, reverseAlign, maxBacktracks,
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triasteran
parents:
diff changeset
401 tryHard, valAlign, allValAligns, suppressAlign, best,
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
402 strata, offrate, seed, snpphred, snpfrac, keepends,
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triasteran
parents:
diff changeset
403 output_unmapped_reads, output_suppressed_reads,
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
404 quality_score_encoding )
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triasteran
parents:
diff changeset
405 except ValueError as e:
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triasteran
parents:
diff changeset
406 # clean up temp dir
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triasteran
parents:
diff changeset
407 if os.path.exists( tmp_index_dir ):
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triasteran
parents:
diff changeset
408 shutil.rmtree( tmp_index_dir )
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triasteran
parents:
diff changeset
409 stop_err( 'Something is wrong with the alignment parameters and the alignment could not be run\n' + str( e ) )
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triasteran
parents:
diff changeset
410 try:
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triasteran
parents:
diff changeset
411 # have to nest try-except in try-finally to handle 2.4
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triasteran
parents:
diff changeset
412 try:
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triasteran
parents:
diff changeset
413 # prepare actual mapping commands
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parents:
diff changeset
414 if options.paired == 'paired':
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triasteran
parents:
diff changeset
415 cmd2 = 'bowtie %s %s -1 %s -2 %s > %s' % ( aligning_cmds, ref_file_name, options.input1, options.input2, options.output )
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
416 else:
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triasteran
parents:
diff changeset
417 cmd2 = 'bowtie %s %s %s > %s' % ( aligning_cmds, ref_file_name, options.input1, options.output )
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
418 # align
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triasteran
parents:
diff changeset
419 tmp = tempfile.NamedTemporaryFile( dir=tmp_index_dir ).name
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triasteran
parents:
diff changeset
420 with open(tmp, 'w') as tmp_stderr:
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triasteran
parents:
diff changeset
421 returncode = subprocess.call(args=cmd2, shell=True, cwd=tmp_index_dir, stderr=tmp_stderr.fileno())
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
422 # get stderr, allowing for case where it's very large
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triasteran
parents:
diff changeset
423 stderr = ''
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triasteran
parents:
diff changeset
424 buffsize = 1048576
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triasteran
parents:
diff changeset
425 with open(tmp, 'r') as tmp_stderr:
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triasteran
parents:
diff changeset
426 try:
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triasteran
parents:
diff changeset
427 while True:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
428 stderr += tmp_stderr.read(buffsize)
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
429 if not stderr or len(stderr) % buffsize != 0:
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triasteran
parents:
diff changeset
430 break
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triasteran
parents:
diff changeset
431 except OverflowError:
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triasteran
parents:
diff changeset
432 pass
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triasteran
parents:
diff changeset
433 if returncode != 0:
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triasteran
parents:
diff changeset
434 raise Exception(stderr)
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
435 elif options.output_mapping_stats is not None:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
436 # Write stderr (containing the mapping statistics) to a named file
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
437 with open(options.output_mapping_stats, 'w') as mapping_stats:
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triasteran
parents:
diff changeset
438 mapping_stats.write( stderr )
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triasteran
parents:
diff changeset
439 # get suppressed and unmapped reads output files in place if appropriate
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triasteran
parents:
diff changeset
440 if options.paired == 'paired' and tmp_suppressed_file_name and \
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
441 options.output_suppressed_reads_l and options.output_suppressed_reads_r:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
442 try:
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triasteran
parents:
diff changeset
443 left = tmp_suppressed_file_name.replace( '.fastq', '_1.fastq' )
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
444 right = tmp_suppressed_file_name.replace( '.fastq', '_1.fastq' )
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
445 shutil.move( left, options.output_suppressed_reads_l )
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
446 shutil.move( right, options.output_suppressed_reads_r )
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
447 except Exception as e:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
448 sys.stdout.write( 'Error producing the suppressed output file.\n' )
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
449 if options.paired == 'paired' and tmp_unmapped_file_name and \
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
450 options.output_unmapped_reads_l and options.output_unmapped_reads_r:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
451 try:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
452 left = tmp_unmapped_file_name.replace( '.fastq', '_1.fastq' )
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
453 right = tmp_unmapped_file_name.replace( '.fastq', '_2.fastq' )
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
454 shutil.move( left, options.output_unmapped_reads_l )
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
455 shutil.move( right, options.output_unmapped_reads_r )
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
456 except Exception as e:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
457 sys.stdout.write( 'Error producing the unmapped output file.\n' )
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
458 # check that there are results in the output file
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triasteran
parents:
diff changeset
459 if os.path.getsize( options.output ) == 0:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
460 raise Exception('The output file is empty, there may be an error with your input file or settings.')
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
461 except Exception as e:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
462 stop_err( 'Error aligning sequence. ' + str( e ) )
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
463 finally:
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
464 # clean up temp dir
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
465 if os.path.exists( tmp_index_dir ):
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
466 shutil.rmtree( tmp_index_dir )
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
467 stdout += 'Sequence file aligned.\n'
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
468 sys.stdout.write( stdout )
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
469
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
470
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
471 if __name__ == "__main__":
7e3bd576aa7c Uploaded
triasteran
parents:
diff changeset
472 __main__()