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1 #!/usr/bin/perl -w
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2
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3 #Margaret Antonio 16.01.13
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4
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5 #DESCRIPTION: Takes two aggregate.pl outputs and compares them using mean difference, pval for each
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6 #gene. Can compare, for example, 19F in glucose and TIGR4 in glucose.
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7 #DIFFERENT GENOMES (ie. diff. strains).
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8 #Requires CONVERSION FILE
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9
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10 #USAGE: perl compStrains.pl -c <conversion.csv> <options>
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11 #[<aggregateFile1.csv aggregateFile2.csv> OR -indir <indir/>]
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12
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13 use Data::Dumper;
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14 use strict;
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15 use Getopt::Long;
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16 use warnings;
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17 use File::Path;
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18 use File::Basename;
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19 use Statistics::Distributions;
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20
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21 #ASSIGN INPUTS TO VARIABLES USING FLAGS
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22 our ($input1,$input2,$out,$sortkey,$round,$l1,$l2,$cfile);
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23 GetOptions(
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24 'h' => \$h,
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25 'o:s' =>\$out,
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26 's:i' => \$sortkey,
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27 'r:i'=> \$round,
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28 'l1:s'=> \$l1,
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29 'l2:s'=> \$l2,
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30 'c:s'=> \$cfile,
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31 'input1:s'=>\$input1,
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32 'input2:s'=>\$input2
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33 );
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34
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35 sub print_usage() {
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36 print "\n";
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37 print "\n##################################################################\n";
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38 print "compStrains.pl: compare genes from a tn-seq experiment\n";
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39 print "\tfor two DIFFERENT strains/genomes using aggregate files\n";
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40
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41 print "\nDESCRIPTION: Takes two aggregate.pl outputs and compares them by\n";
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42 print "calculating the difference in mean fitness.\n";
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43
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44 print "Example: two strains tested under same condition.\n";
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45 print "Note: For same strains (genomes), use compGenes.pl\n";
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46
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47 print "\nUSAGE:\n";
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48 print "perl compStrains.pl -c conversion.csv -d inputs/\n";
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49
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50 print "\nREQUIRED:\n";
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51 print " -d\tDirectory containing all input files (files from\n";
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52 print " \taggregate fitness script)\n";
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53 print " \tOR\n";
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54 print " \tIn the command line (without a flag), input the name(s) of\n";
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55 print " \ttwo files containing aggregate gene fitness values. \n\n";
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56 print " -c\tConversion file: two columns with homologs for both organisms\n";
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57
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58 print "\nOPTIONAL:\n";
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59 print " -h\tPrints usage and exits program\n";
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60 print " -o\tOutput file for comparison data. Default: label1label2.csv\n";
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61 print " -s\tSort output by this index of the file (indices begin at 0).\n";
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62 print " \tDefault: by mean\n";
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63 print " -r\tRound final output numbers to this number of decimals\n";
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64 print " -l\tLabels for input files. Default: filenames\n";
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65 print " \tTwo strings, comma separated (i.e. -l expt1,expt2).\n";
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66 print " \tOrder should match file order.\n";
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67 print " \n~~~~Always check that file paths are correctly specified~~~~\n";
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68 print "\n##################################################################\n";
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69 }
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70
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71 #THE @files ARRAY WILL CONTAIN INPUT FILE NAMES
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72 my @files=($input1,$input2)
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73
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74 #GET LABELS:
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75 my @labels = ($l1,$l2);
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76
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77
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78 #CHECK IF REQ. VARIABLES WERE DEFINED USING FLAGS. IF NOT THEN USE DEFAULT VALUES
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79
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80 if (!$out) {$out="comp.".$labels[0].$labels[1].".csv"}
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81 if (!$round){$round='%.4f'}
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82
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83 #OPEN INPUTTED AGGREGATE GENE FILES AND STORE THEIR CONTENTS INTO TWO HASHES
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84 #FILE1 GOES INTO HASH %ONE AND FILE2 GOES INTO HASH %TWO.
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85
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86 #FILE1 OPENING ---> %one WHERE KEY:VALUE IS GENE_ID:(GENE_ID,INSERTIONS,MEAN,ETC.)
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87 my @header;
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88 my %one;
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89
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90 open (F1,'<',$files[0]);
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91
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92 #STORE COLUMN NAMES (FIRST LINE OF FILE1) FOR HEADER AND APPEND LABELS
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93 my $head=<F1>; #the header in the file
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94 my @cols=split(',',$head);
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95 @cols=@cols[0,1,2,3,4,5,6]; #get rid of blank columns
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96 for (my $j=0;$j<scalar @cols;$j++){
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97 $cols[$j]=$cols[$j].'-'.$labels[0]; #mark each column name with file it comes from
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98 }
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99 push (@header,@cols);
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100
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101 while (my $line=<F1>){
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102 chomp $line;
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103 my @info=split(",",$line);
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104 #Only keep the first 7 columns (Ones about blanks aren't needed for comparisons)
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105 @info=@info[0,1,2,3,4,5,6];
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106 #Sometimes genes that don't have a gene name can't be blank, so fill with NA
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107 if (!$info[5]){
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108 $info[5]="NA";
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109 }
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110 #If there are no insertions in the column "total", then make it =0 rather than blank
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111 if (!$info[6]){
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112 $info[6]=0;
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113 }
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114 $one{$info[0]}=\@info;
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115 }
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116 close F1;
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117
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118 #FILE2 OPENING ---> %two WHERE KEY:VALUE IS GENE_ID:(GENE_ID,INSERTIONS,MEAN,ETC.)
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119
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120 my %two;
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121 open (F2,'<',$files[1]);
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122
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123 #STORE COLUMN NAMES (FIRST LINE OF FILE2) FOR HEADER AND APPEND LABELS
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124 $head=<F2>; #the header in the file
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125 @cols=split(',',$head);
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126 @cols=@cols[0,1,2,3,4,5,6]; #get rid of blank columns
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127 for (my $j=0;$j<scalar @cols;$j++){
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128 $cols[$j]=$cols[$j].'-'.$labels[1]; #mark each column name with file it comes from
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129 }
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130 push (@header,@cols);
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131
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132 while (my $line=<F2>){
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133 chomp $line;
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134 my @info=split(",",$line);
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135 @info=@info[0,1,2,3,4,5,6];
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136 if (!$info[5]){
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137 $info[5]="NA";
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138 }
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139 if (!$info[6]){
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140 $info[6]=0;
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141 }
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142 $two{$info[0]}=\@info;
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143 }
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144 close F2;
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145
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146
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147 #READ CONVERSION FILE INTO ARRAY.
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148 #Conversion file must have strain 1 for file 1 in column 1 (index 0) and
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149 #strain 2 for file 2 in column 2 (index 1)
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150 #conversion file must be tab delimited with no NA fields
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151 #If homologs (exist then take info from hashes (%one and %two) by referring to gene_id in KEY
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152
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153 my @all; #store all homologs in this hash
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154 open (CONV,'<',$cfile);
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155 while (my $line=<CONV>){
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156 chomp $line;
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157 my @genes=split("\t",$line); #Array @genes will contain two genes (SP_0000,SPT_0000)
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158 if (scalar @genes==2 and $genes[0] ne "" and $genes[1] ne ""){
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159 my @info;
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160 my @oneArray=@{$one{$genes[0]}};
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161 my @twoArray=@{$two{$genes[1]}};
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162 push (@info,@oneArray,@twoArray);
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163 my $diff=sprintf("$round",($info[1]-$info[8]));
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164 my $total1=$info[6];
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165 my $total2=$info[13];
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166 my $sd1=$info[3];
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167 my $se1=$info[4];
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168 my $sd2=$info[10];
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169 my $se2=$info[11];
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170 my $df=$total1+$total2-2;
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171 my $tdist;
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172 my $pval;
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173 #TDIST, PVAL calculations with fail if standard dev, error, or counts are not real numbers
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174 #or if 0 ends up in denominator
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175 if ($se1 eq "X" or $se2 eq "X" or $sd1 eq "X" or $sd2 eq "X" or $total1==0 or $total2==0 or $sd1==0 or $sd2==0){
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176 ($tdist,$pval)=("NA","NA");
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177 }
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178 else{
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179 $tdist=sqrt((($diff)/(sqrt((($sd1**2)/$total1)+(($sd2**2)/$total2))))**2);
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180 $pval=Statistics::Distributions::tprob($df,$tdist);
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181 }
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182 push (@info,$diff,$df,$tdist,$pval);
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183 push (@all,\@info);
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184 }
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185 }
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186 close CONV;
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187
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188 #SORT THE HOMOLOGS BY THE SORTKEY OR BY DEFAULT DIFFERENCE IN MEAN FITNESSES
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189 if (!$sortkey){
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190 $sortkey=14; #for mean difference
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191 }
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192 my @sorted = sort { $b->[$sortkey] <=> $a->[$sortkey] } @all;
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193
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194 #FINISH THE HEADER BY ADDING COLUMN NAMES FOR MEAN-DIFF, DOF, TDIST, AND PVALUE
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195 my $field="MeanDiff(".$labels[0].'.'.$labels[1].")";
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196 push (@header,$field,"DOF","TDIST","PVALUE");
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197
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198 #PRINT MATCHED HOMOLOG INFORMATION INTO A SINGLE OUTPUT FILE
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199 open OUT, '>',"$out";
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200 print OUT (join(',',@header),"\n");
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201 foreach (@sorted){
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202 my @woo=@{$_};
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203 print OUT join(',',@woo),"\n";
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204 }
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205
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206 close OUT;
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207
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208
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