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1 #!/usr/bin/perl
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2 my $SrcFolder="/home/galaxy/galaxy-dist/tools/SpliceTrap.0.90.1/bin";
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3 # Author: wuj@cshl.edu
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4 use strict;
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5 use Getopt::Long;
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6 ####################
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7 use Cwd;
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8 my $PROG = $0;
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9 my $CUR_DIR = Cwd::abs_path(Cwd::cwd());
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10 my $PROG_ABS_PATH = Cwd::abs_path($PROG);
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11 #my $SrcFolder=`dirname $PROG_ABS_PATH`;
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12 #chomp($SrcFolder);
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13 #my %config=do "$ENV{HOME}/.SpliceTrap.pl.ini";
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14 #my $SrcFolder=$config{SrcFolder};
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15
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16 my @programs = ('R','echo','cat','bash','perl','ln','mkdir','paste','grep','sort','basename','awk','wc','mv','cd','rm','split','head' );
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17 foreach my $program (@programs)
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18 {
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19 die ("CHECK: $program not found\n") if(system("hash $program >/dev/null"));
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20
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21 }
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22
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23 ####################
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24 my $MapSoftware="bowtie";
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25 my $DatabasePrefix="hg18";
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26 my $ReadFileFormat="";
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27 my $ReadFile1Name="";
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28 my $ReadFile2Name="";
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29 my $CutoffLevel="M";
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30 my $Outputfolder=$CUR_DIR;
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31 my $OutputPrefix="Result";
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32 #my $CutoffOnly=0;
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33 my $ReadSize=36;
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34 my $JunctionCut=5;
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35 my $onGalaxy="";
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36 my $BowtieThreads=1;
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37 my $noIRMstr="";
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38 my $noIRM = 0;
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39
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40 GetOptions (
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41 "m:s"=>\$MapSoftware,
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42 "d:s"=>\$DatabasePrefix,
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43 # "f:s"=>\$ReadFileFormat,
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44 "1:s"=>\$ReadFile1Name,
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45 "2:s"=>\$ReadFile2Name,
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46 "c:s"=>\$CutoffLevel,
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47 "outdir:s"=>\$Outputfolder,
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48 "o:s"=>\$OutputPrefix,
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49 "j:i"=>\$JunctionCut,
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50 "s:i"=>\$ReadSize,
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51 "p:i"=>\$BowtieThreads,
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52 "noIRM|noirm"=>\$noIRM,
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53 "g:s"=>\$onGalaxy
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54 # "local:s"=>\$local,
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55 # "rerun"=>\$CutoffOnly
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56 );
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57 #-O for galaxy output
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58
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59
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60 my $InputParaDes=" Usage of the script:
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61 -m Mapping software: [bowtie]/rmap
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62 -d Database prefix: [hg18]/mm9/rn4/userdefined
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63 -1 Read File 1
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64 -2 Read File 2
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65 -c Cutoff Level:H/[M]/L
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66 Means High, Middle or Low
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67 -j Junction reads requirement per junction for each exon-isoform [5]
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68 -o Output prefix {Result}
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69 -s Read Size [36]
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70 --outdir Output folder [./]
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71 -p Bowtie parameter, threads number, only use this when you don't use qsub [1]
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72 --noIRM Skip the IRM correction step
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73
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74 This is a quick help, please refer to the README file for details.
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75 ";
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76
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77 if($ReadFile2Name eq "")
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78 {
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79 $ReadFile2Name = $ReadFile1Name;
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80 #trigger singled end mode
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81 }
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82
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83 if($ReadFile1Name eq "" or $ReadFile2Name eq "" )
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84 {
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85 print $InputParaDes;
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86 exit;
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87 }
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88
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89 if($BowtieThreads < 1)
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90 {
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91 print $InputParaDes;
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92 exit;
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93 }
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94
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95 if (! -e "$SrcFolder/../db/$DatabasePrefix/parallel")
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96 {
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97 print "CHECK: Error, the database you specified is not properly installed.\n";
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98 #print $InputParaDes;
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99 exit;
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100
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101 }
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102
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103 if($CutoffLevel ne "H" and $CutoffLevel ne "M" and $CutoffLevel ne "L")
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104 {
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105 print $InputParaDes;
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106 exit;
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107 }
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108
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109 $ReadFile1Name = Cwd::abs_path($ReadFile1Name);
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110 $ReadFile2Name = Cwd::abs_path($ReadFile2Name);
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111
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112 #check the files
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113 open(check,$ReadFile1Name) or die ("CHECK: Error when opening $ReadFile1Name\n");
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114 my $checkoneline = <check>;
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115 if(substr($checkoneline,0,1) eq ">")
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116 {
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117 $ReadFileFormat = "fasta";
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118 }
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119 elsif(substr($checkoneline,0,1) eq "@")
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120 {
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121 $ReadFileFormat = "fastq";
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122 }
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123 else
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124 {
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125 die("CHECK: ERROR:Please check $ReadFile1Name\n");
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126 }
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127 close(check);
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128
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129 open(check,$ReadFile2Name) or die ("CHECK: Error when opening $ReadFile2Name\n");
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130 my $checkoneline = <check>;
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131 if(substr($checkoneline,0,1) eq ">")
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132 {
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133 die("CHECK: $ReadFile2Name has a different format as $ReadFile1Name\n") if ($ReadFileFormat ne "fasta");
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134 }
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135 elsif(substr($checkoneline,0,1) eq "@")
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136 {
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137 die("CHECK: $ReadFile2Name has a different format as $ReadFile1Name\n") if ($ReadFileFormat ne "fastq");
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138 }
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139 else
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140 {
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141 die("CHECK: ERROR:Please check $ReadFile2Name\n");
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142 }
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143 close(check);
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144
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145 $Outputfolder= Cwd::abs_path($Outputfolder);
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146 if($Outputfolder eq "/tmp")
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147 {
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148 while(-e $Outputfolder)
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149 {
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150 my $random_foldername = random_sessid();
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151 $Outputfolder = "/tmp/".$random_foldername;
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152 }
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153 }
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154
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155
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156 if(! -e $Outputfolder)
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157 {
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158 mkdir $Outputfolder or die "CHECK: cannot mkdir $Outputfolder\n";
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159 }
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160 if(! -d $Outputfolder)
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161 {
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162 die "CHECK: $Outputfolder is not a folder\n";
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163 }
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164
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165 if($MapSoftware eq "bowtie")
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166 {
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167 print "CHECK: whether bowtie installed and in PATH\n";
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168 my $bowtiechecker=`bowtie --version`;
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169 if($bowtiechecker ne "")
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170 {
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171 print "CHECK: bowtie found, information below:\n";
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172 print $bowtiechecker,"\n";
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173 }
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174 else
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175 {
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176 die "CHECK: No bowtie found in PATH, EXIT!\n";
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177 }
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178 }
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179 elsif($MapSoftware eq "rmap")
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180 {
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181 print "CHECK: checking rmap...\n";
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182 if(system("type rmap &>/dev/null") ==0 )
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183 {
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184 print "CHECK: rmap found, continue\n";
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185 }
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186 else
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187 {
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188 die "CHECK: No rmap found in PATH, EXIT!\n";
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189 }
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190 }
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191 else
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192 {
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193 die "CHECK: option -m only takes rmap or bowtie as inputs\n";
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194 }
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195
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196 if($ReadSize == 0)
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197 {
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198 die "CHECK: Please check option -s Read size\n";
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199 }
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200
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201 if($noIRM)
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202 {
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203 $noIRMstr= "noirm";
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204 }
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205
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206 #write more checks later
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207 print "PARAMETERS:\tMapping software: ",$MapSoftware,"\n";
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208 print "PARAMETERS:\tDatabase prefix: ",$DatabasePrefix,"\n";
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209 print "PARAMETERS:\tRead end 1: ",$ReadFile1Name,"\n";
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210 print "PARAMETERS:\tRead end 2: ",$ReadFile2Name,"\n" if($ReadFile2Name ne $ReadFile1Name);
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211 print "PARAMETERS:\tCutoff level: ",$CutoffLevel,"\n";
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212 print "PARAMETERS:\tJunction reads.min:",$JunctionCut,"\n";
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213 print "PARAMETERS:\tOutput folder: ",$Outputfolder,"\n";
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214 print "PARAMETERS:\tOutput prefix: ",$OutputPrefix,"\n";
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215 print "PARAMETERS:\tRead size: ",$ReadSize,"\n";
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216 print "PARAMETERS:\tBowtie threads #: ",$BowtieThreads,"\n";
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217 print "PARAMETERS:\tNo IRM.\n" if ($noIRM);
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218
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219 if($MapSoftware eq "bowtie")
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220 {
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221 print "=================STAGE 1 MAPPING===================\n";
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222 system("bash $SrcFolder/mapping_bowtie.sh $ReadFile1Name $ReadFileFormat $DatabasePrefix $Outputfolder $SrcFolder $BowtieThreads");
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223 system("bash $SrcFolder/mapping_bowtie.sh $ReadFile2Name $ReadFileFormat $DatabasePrefix $Outputfolder $SrcFolder $BowtieThreads") if($ReadFile2Name ne $ReadFile1Name);
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224 print "=================STAGE 2 ESTIMATION================\n";
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225
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226 system("bash $SrcFolder/batch_para_cov10p_fit.sh $ReadFile1Name $ReadFile2Name $OutputPrefix $ReadSize $DatabasePrefix $Outputfolder $SrcFolder $noIRMstr") ;
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227 print "=================STAGE 3 CUTOFF====================\n";
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228 system("bash $SrcFolder/apply_cutoff.sh $OutputPrefix $CutoffLevel $Outputfolder $JunctionCut $SrcFolder $noIRMstr");
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229
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230
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231 }
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232
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233 if($MapSoftware eq "rmap")
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234 {
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235 print "=================STAGE 1 MAPPING===================\n";
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236
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237 system("bash $SrcFolder/mapping_rmap.sh $ReadFile1Name $ReadFileFormat $DatabasePrefix $Outputfolder $SrcFolder") ;
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238 system("bash $SrcFolder/mapping_rmap.sh $ReadFile2Name $ReadFileFormat $DatabasePrefix $Outputfolder $SrcFolder") if($ReadFile2Name ne $ReadFile1Name);
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239 print "=================STAGE 2 ESTIMATION================\n";
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240
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241 system("bash $SrcFolder/batch_para_cov10p_fit.sh $ReadFile1Name $ReadFile2Name $OutputPrefix $ReadSize $DatabasePrefix $Outputfolder $SrcFolder $noIRMstr") ;
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242 print "=================STAGE 3 CUTOFF====================\n";
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243 system("bash $SrcFolder/apply_cutoff.sh $OutputPrefix $CutoffLevel $Outputfolder $JunctionCut $SrcFolder $noIRMstr");
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244
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245
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246 }
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247
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248 print "============ALL DONE, OUTPUTFILE:$OutputPrefix.txt\n";
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249
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250 if($onGalaxy ne "")
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251 {
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252 system("grep -v na $Outputfolder/$OutputPrefix.txt >$onGalaxy");
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253 }
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254
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255 sub random_sessid
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256 {
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257 #my @chars = (0..9,a..z,A..Z);
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258 my @chars = ('a'..'z','A'..'Z');
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259 my $len = 10;
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260 my $string = join '', map {$chars[rand(@chars)]} (1..$len);
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261 return $string;
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262 }
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263
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