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1 #!/usr/bin/perl
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2 # Author: wuj@cshl.edu
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3 # Modified: Baekdoo Kim (baegi7942@gmail.com)
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4 use strict;
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5 use Getopt::Long;
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6 use Data::Dumper;
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7 ####################
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8 use Cwd;
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9 my $PROG = $0;
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10 my $CUR_DIR = Cwd::abs_path(Cwd::cwd());
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11 my $PROG_ABS_PATH = Cwd::abs_path($PROG);
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12 #my $SrcFolder=`dirname $PROG_ABS_PATH`;
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13 #chomp($SrcFolder);
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14 #my %config=do "$ENV{HOME}/.SpliceTrap.pl.ini";
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15 #my $SrcFolder=$config{SrcFolder};
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16
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17 my @programs = ('R','echo','cat','bash','perl','ln','mkdir','paste','grep','sort','basename','awk','wc','mv','cd','rm','split','head' );
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18 foreach my $program (@programs)
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19 {
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20 die ("CHECK: $program not found\n") if(system("hash $program >/dev/null"));
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21
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22 }
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23
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24 ####################
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25 my $SrcFolder="";
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26 my $MapSoftware="bowtie";
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27 my $DatabasePrefix="hg38";
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28 my $ReadFileFormat="";
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29 my $ReadFile1Name="";
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30 my $ReadFile2Name="";
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31 my $CutoffLevel="M";
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32 my $Outputfolder=$CUR_DIR;
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33 my $OutputPrefix="Result";
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34 #my $CutoffOnly=0;
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35 my $ReadSize=36;
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36 my $JunctionCut=5;
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37 my $onGalaxy_raw="";
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38 my $onGalaxy_txt="";
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39 my $BowtieThreads=1;
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40 my $noIRMstr="";
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41 my $noIRM = 0;
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42
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43 my $num_args = $#ARGV;
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44 $onGalaxy_raw = $ARGV[$num_args-1];
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45 $onGalaxy_txt = $ARGV[$num_args];
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46
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47 GetOptions (
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48 "l:s"=>\$SrcFolder,
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49 "m:s"=>\$MapSoftware,
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50 "d:s"=>\$DatabasePrefix,
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51 # "f:s"=>\$ReadFileFormat,
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52 "1:s"=>\$ReadFile1Name,
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53 "2:s"=>\$ReadFile2Name,
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54 "c:s"=>\$CutoffLevel,
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55 "outdir:s"=>\$Outputfolder,
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56 "o:s"=>\$OutputPrefix,
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57 "j:i"=>\$JunctionCut,
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58 "s:i"=>\$ReadSize,
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59 "p:i"=>\$BowtieThreads,
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60 "noIRM|noirm"=>\$noIRM
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61 # "local:s"=>\$local,
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62 # "rerun"=>\$CutoffOnly
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63 );
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64 #-O for galaxy output
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65
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66
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67 my $InputParaDes=" Usage of the script:
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68 -l Base Location (required)
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69 -m Mapping software: [bowtie]/rmap
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70 -d Database prefix: [hg18]/mm9/rn4/userdefined
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71 -1 Read File 1
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72 -2 Read File 2
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73 -c Cutoff Level:H/[M]/L
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74 Means High, Middle or Low
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75 -j Junction reads requirement per junction for each exon-isoform [5]
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76 -o Output prefix {Result}
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77 -s Read Size [36]
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78 --outdir Output folder [./]
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79 -p Bowtie parameter, threads number, only use this when you don't use qsub [1]
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80 --noIRM Skip the IRM correction step
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81
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82 This is a quick help, please refer to the README file for details.
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83 ";
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84
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85
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86 if($SrcFolder eq "") {
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87 print "[CHECK] - Please provide the location of the script (option '-l')\n\n";
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88 exit;
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89 }
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90
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91 if($ReadFile2Name eq "")
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92 {
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93 $ReadFile2Name = $ReadFile1Name;
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94 #trigger singled end mode
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95 }
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96
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97 if($ReadFile1Name eq "" or $ReadFile2Name eq "" )
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98 {
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99 print $InputParaDes;
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100 exit;
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101 }
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102
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103 if($BowtieThreads < 1)
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104 {
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105 print $InputParaDes;
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106 exit;
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107 }
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108
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109 if (! -e "$SrcFolder/db/$DatabasePrefix/parallel")
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110 {
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111 print "CHECK: Error, the database you specified is not properly installed.\n";
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112 #print $InputParaDes;
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113 exit;
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114
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115 }
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116
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117 if($CutoffLevel ne "H" and $CutoffLevel ne "M" and $CutoffLevel ne "L")
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118 {
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119 print $InputParaDes;
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120 exit;
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121 }
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122
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123
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124 $ReadFile1Name = Cwd::abs_path($ReadFile1Name);
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125 $ReadFile2Name = Cwd::abs_path($ReadFile2Name);
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126
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127 #check the files
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128 open(check,$ReadFile1Name) or die ("CHECK: Error when opening $ReadFile1Name\n");
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129 my $checkoneline = <check>;
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130 if(substr($checkoneline,0,1) eq ">")
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131 {
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132 $ReadFileFormat = "fasta";
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133 }
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134 elsif(substr($checkoneline,0,1) eq "@")
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135 {
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136 $ReadFileFormat = "fastq";
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137 }
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138 else
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139 {
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140 die("CHECK: ERROR:Please check $ReadFile1Name\n");
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141 }
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142 close(check);
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143
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144 open(check,$ReadFile2Name) or die ("CHECK: Error when opening $ReadFile2Name\n");
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145 my $checkoneline = <check>;
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146 if(substr($checkoneline,0,1) eq ">")
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147 {
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148 die("CHECK: $ReadFile2Name has a different format as $ReadFile1Name\n") if ($ReadFileFormat ne "fasta");
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149 }
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150 elsif(substr($checkoneline,0,1) eq "@")
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151 {
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152 die("CHECK: $ReadFile2Name has a different format as $ReadFile1Name\n") if ($ReadFileFormat ne "fastq");
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153 }
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154 else
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155 {
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156 die("CHECK: ERROR:Please check $ReadFile2Name\n");
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157 }
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158 close(check);
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159
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160 $Outputfolder= Cwd::abs_path($Outputfolder);
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161 if($Outputfolder eq "/tmp")
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162 {
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163 while(-e $Outputfolder)
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164 {
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165 my $random_foldername = random_sessid();
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166 $Outputfolder = "/tmp/".$random_foldername;
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167 }
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168 }
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169
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170
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171 if(! -e $Outputfolder)
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172 {
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173 mkdir $Outputfolder or die "CHECK: cannot mkdir $Outputfolder\n";
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174 }
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175 if(! -d $Outputfolder)
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176 {
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177 die "CHECK: $Outputfolder is not a folder\n";
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178 }
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179
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180 if($MapSoftware eq "bowtie")
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181 {
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182 print "CHECK: whether bowtie installed and in PATH\n";
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183 my $bowtiechecker=`bowtie --version`;
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184 if($bowtiechecker ne "")
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185 {
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186 print "CHECK: bowtie found, information below:\n";
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187 print $bowtiechecker,"\n";
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188 }
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189 else
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190 {
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191 die "CHECK: No bowtie found in PATH, EXIT!\n";
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192 }
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193 }
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194 elsif($MapSoftware eq "rmap")
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195 {
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196 print "CHECK: checking rmap...\n";
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197 if(system("type rmap &>/dev/null") ==0 )
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198 {
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199 print "CHECK: rmap found, continue\n";
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200 }
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201 else
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202 {
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203 die "CHECK: No rmap found in PATH, EXIT!\n";
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204 }
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205 }
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206 else
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207 {
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208 die "CHECK: option -m only takes rmap or bowtie as inputs\n";
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209 }
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210
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211 if($ReadSize == 0)
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212 {
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213 die "CHECK: Please check option -s Read size\n";
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214 }
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215
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216 if($noIRM)
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217 {
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218 $noIRMstr= "noirm";
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219 }
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220
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221 #write more checks later
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222 print "PARAMETERS:\tMapping software: ",$MapSoftware,"\n";
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223 print "PARAMETERS:\tDatabase prefix: ",$DatabasePrefix,"\n";
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224 print "PARAMETERS:\tRead end 1: ",$ReadFile1Name,"\n";
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225 print "PARAMETERS:\tRead end 2: ",$ReadFile2Name,"\n" if($ReadFile2Name ne $ReadFile1Name);
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226 print "PARAMETERS:\tGalaxy_raw: ",$onGalaxy_raw,"\n"; #if($onGalaxy_raw ne "");
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227 print "PARAMETERS:\tGalaxy_txt: ",$onGalaxy_txt,"\n"; #if($onGalaxy_txt ne "");
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228 print "PARAMETERS:\tCutoff level: ",$CutoffLevel,"\n";
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229 print "PARAMETERS:\tJunction reads.min:",$JunctionCut,"\n";
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230 print "PARAMETERS:\tOutput folder: ",$Outputfolder,"\n";
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231 print "PARAMETERS:\tOutput prefix: ",$OutputPrefix,"\n";
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232 print "PARAMETERS:\tRead size: ",$ReadSize,"\n";
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233 print "PARAMETERS:\tBowtie threads #: ",$BowtieThreads,"\n";
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234 print "PARAMETERS:\tNo IRM.\n" if ($noIRM);
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235
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236 if($MapSoftware eq "bowtie")
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237 {
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238 print "=================STAGE 1 MAPPING===================\n";
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239 system("bash $SrcFolder/bin/mapping_bowtie.sh $ReadFile1Name $ReadFileFormat $DatabasePrefix $Outputfolder $SrcFolder $BowtieThreads");
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240 system("bash $SrcFolder/bin/mapping_bowtie.sh $ReadFile2Name $ReadFileFormat $DatabasePrefix $Outputfolder $SrcFolder $BowtieThreads") if($ReadFile2Name ne $ReadFile1Name);
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241 print "=================STAGE 2 ESTIMATION================\n";
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242 # ratio, log, nums
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243 system("bash $SrcFolder/bin/batch_para_cov10p_fit.sh $ReadFile1Name $ReadFile2Name $OutputPrefix $ReadSize $DatabasePrefix $Outputfolder $SrcFolder $noIRMstr") ;
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244 print "=================STAGE 3 CUTOFF====================\n";
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245 # raw
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246 system("bash $SrcFolder/bin/apply_cutoff.sh $OutputPrefix $CutoffLevel $Outputfolder $JunctionCut $SrcFolder $noIRMstr");
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247
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248
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249 }
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250
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251 if($MapSoftware eq "rmap")
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252 {
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253 print "=================STAGE 1 MAPPING===================\n";
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254
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255 system("bash $SrcFolder/bin/mapping_rmap.sh $ReadFile1Name $ReadFileFormat $DatabasePrefix $Outputfolder $SrcFolder") ;
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256 system("bash $SrcFolder/bin/mapping_rmap.sh $ReadFile2Name $ReadFileFormat $DatabasePrefix $Outputfolder $SrcFolder") if($ReadFile2Name ne $ReadFile1Name);
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257 print "=================STAGE 2 ESTIMATION================\n";
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258
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259 system("bash $SrcFolder/bin/batch_para_cov10p_fit.sh $ReadFile1Name $ReadFile2Name $OutputPrefix $ReadSize $DatabasePrefix $Outputfolder $SrcFolder $noIRMstr") ;
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260 print "=================STAGE 3 CUTOFF====================\n";
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261 system("bash $SrcFolder/bin/apply_cutoff.sh $OutputPrefix $CutoffLevel $Outputfolder $JunctionCut $SrcFolder $noIRMstr");
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262
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263
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264 }
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265
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266 #print "============ALL DONE, OUTPUTFILE:$OutputPrefix.txt\n";
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267
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268 if($onGalaxy_raw ne "" && $onGalaxy_txt ne "")
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269 {
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270 print "OUTPUTFILE:$OutputPrefix.raw\n";
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271 system("grep -v na $Outputfolder/$OutputPrefix.raw >$onGalaxy_raw");
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272 print "OUTPUTFILE:$OutputPrefix.txt\n";
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273 system("grep -v na $Outputfolder/$OutputPrefix.txt >$onGalaxy_txt");
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274 }
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275
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276 print "============Clean up\n";
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277 system("rm -r $Outputfolder/$OutputPrefix.*");
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278
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279 sub random_sessid {
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280 #my @chars = (0..9,a..z,A..Z);
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281 my @chars = ('a'..'z','A'..'Z');
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282 my $len = 10;
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283 my $string = join '', map {$chars[rand(@chars)]} (1..$len);
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284 return $string;
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285 }
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