Mercurial > repos > yhoogstrate > edger_with_design_matrix
annotate edgeR_Differential_Gene_Expression.xml @ 2:ec951a5017f8 draft
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit a6cf3ec153ca4a3846258a223d287ca125eea7be
author | yhoogstrate |
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date | Tue, 01 Sep 2015 09:15:07 -0400 |
parents | a4a4c88783ea |
children | 12fb0d4b1e93 |
rev | line source |
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a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
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1 <?xml version="1.0" encoding="UTF-8"?> |
2
ec951a5017f8
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit a6cf3ec153ca4a3846258a223d287ca125eea7be
yhoogstrate
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2 <tool id="edger_dge" name="edgeR: Differential Gene(Expression) Analysis" version="3.11.0.b"> |
1
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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3 <description>RNA-Seq gene expression analysis using edgeR (R package)</description> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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4 |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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5 <macros> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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6 <import>edgeR_macros.xml</import> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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7 </macros> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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8 |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
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9 <requirements> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
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10 <requirement type="package" version="3.11.0">edger</requirement> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
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11 </requirements> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
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12 |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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13 <stdio> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
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14 <regex match="Error in[^a-z]+contrasts" |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
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15 source="both" |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
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16 level="fatal" |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
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17 description="Have the design- and expression-matrix been swapped?" /> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
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18 <regex match="Execution halted" |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
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19 source="both" |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
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20 level="fatal" /> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
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21 <regex match="Calculating library sizes from column" |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
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22 source="stderr" |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
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23 level="log" /> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
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24 <regex match="During startup - Warning messages" |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
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25 source="stderr" |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
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26 level="log" /> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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27 <regex match="Setting LC_[^ ]+ failed" |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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28 source="stderr" |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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29 level="warning" |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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30 description="LOCALE has not been set correctly" /> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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31 </stdio> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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32 |
2
ec951a5017f8
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit a6cf3ec153ca4a3846258a223d287ca125eea7be
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33 <version_command>echo $(R --version | grep version | grep -v GNU)", EdgeR version" $(R --vanilla --slave -e "library(edgeR) ; cat(sessionInfo()\$otherPkgs\$edgeR\$Version)" 2> /dev/null | grep -v -i "WARNING: ")</version_command> |
1
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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34 |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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35 <command> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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36 R --vanilla --slave -f $R_script '--args |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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37 $expression_matrix |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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38 $design_matrix |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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39 $contrast |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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40 |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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41 $fdr |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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42 |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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43 $output_count_edgeR |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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44 $output_cpm |
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planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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45 |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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46 /dev/null <!-- Calculation of FPKM/RPKM should come here --> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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47 |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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48 #if $output_raw_counts: |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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49 $output_raw_counts |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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50 #else: |
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planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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51 /dev/null |
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52 #end if |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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53 |
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planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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54 #if $output_MDSplot_logFC: |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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55 $output_MDSplot_logFC |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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56 #else: |
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57 /dev/null |
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58 #end if |
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59 |
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planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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60 #if $output_MDSplot_bcv: |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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61 $output_MDSplot_bcv |
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62 #else: |
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63 /dev/null |
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64 #end if |
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65 |
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66 #if $output_BCVplot: |
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planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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67 $output_BCVplot |
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planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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68 #else: |
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69 /dev/null |
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70 #end if |
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71 |
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72 #if $output_MAplot: |
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73 $output_MAplot |
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74 #else: |
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75 /dev/null |
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76 #end if |
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77 |
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78 #if $output_PValue_distribution_plot: |
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79 $output_PValue_distribution_plot |
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80 #else: |
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81 /dev/null |
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82 #end if |
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83 |
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84 #if $output_hierarchical_clustering_plot: |
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85 $output_hierarchical_clustering_plot |
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86 #else: |
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87 /dev/null |
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88 #end if |
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89 |
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90 #if $output_heatmap_plot: |
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91 $output_heatmap_plot |
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92 #else: |
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93 /dev/null |
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94 #end if |
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95 |
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96 #if $output_RData_obj: |
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97 $output_RData_obj |
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98 #else: |
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99 /dev/null |
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100 #end if |
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101 |
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102 $output_format_images |
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103 ' |
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104 </command> |
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105 |
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106 <configfiles> |
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107 <configfile name="R_script"> |
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108 <![CDATA[ |
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109 |
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110 library(limma,quietly=TRUE) ## quietly to avoid unnecessaity stderr messages |
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111 library(edgeR,quietly=TRUE) ## quietly to avoid unnecessaity stderr messages |
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112 library(splines,quietly=TRUE)## quietly to avoid unnecessaity stderr messages |
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113 |
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114 ## Fetch commandline arguments |
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115 args <- commandArgs(trailingOnly = TRUE) |
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116 |
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117 expression_matrix_file <- args[1] |
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118 design_matrix_file <- args[2] |
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119 contrast <- args[3] |
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120 |
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121 fdr <- args[4] |
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122 |
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123 output_count_edgeR <- args[5] |
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124 output_cpm <- args[6] |
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125 |
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126 output_xpkm <- args[7] ##FPKM file - to be implemented |
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127 |
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128 output_raw_counts <- args[8] |
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129 output_MDSplot_logFC <- args[9] |
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130 output_MDSplot_bcv <- args[10] |
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131 output_BCVplot <- args[11] |
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132 output_MAplot <- args[12] |
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133 output_PValue_distribution_plot <- args[13] |
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134 output_hierarchical_clustering_plot <- args[14] |
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135 output_heatmap_plot <- args[15] |
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136 output_RData_obj <- args[16] |
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137 output_format_images <- args[17] |
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138 |
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139 |
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140 ## Obtain read-counts |
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141 expression_matrix <- read.delim(expression_matrix_file,header=T,stringsAsFactors=F,row.names=1,check.names=FALSE,na.strings=c("")) |
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142 design_matrix <- read.delim(design_matrix_file,header=T,stringsAsFactors=F,row.names=1,check.names=FALSE,na.strings=c("")) |
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143 |
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144 colnames(design_matrix) <- make.names(colnames(design_matrix)) |
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145 |
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146 for(i in 1:ncol(design_matrix)) { |
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147 old <- design_matrix[,i] |
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148 design_matrix[,i] <- make.names(design_matrix[,i]) |
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149 if(paste(design_matrix[,i],collapse="\t") != paste(old,collapse="\t")) { |
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150 print("Renaming of factors:") |
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151 print(old) |
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152 print("To:") |
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153 print(design_matrix[,i]) |
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154 } |
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155 ## The following line seems to malfunction the script: |
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156 ##design_matrix[,i] <- as.factor(design_matrix[,i]) |
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157 } |
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158 |
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159 ## 1) In the expression matrix, you only want to have the samples described in the design matrix |
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160 columns <- match(rownames(design_matrix),colnames(expression_matrix)) |
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161 columns <- columns[!is.na(columns)] |
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162 read_counts <- expression_matrix[,columns] |
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163 |
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164 ## 2) In the design matrix, you only want to have samples of which you really have the counts |
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165 columns <- match(colnames(read_counts),rownames(design_matrix)) |
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166 columns <- columns[!is.na(columns)] |
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167 design_matrix <- design_matrix[columns,,drop=FALSE] |
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168 |
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169 ## Filter for HTSeq predifined counts: |
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170 exclude_HTSeq <- c("no_feature","ambiguous","too_low_aQual","not_aligned","alignment_not_unique") |
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171 exclude_DEXSeq <- c("_ambiguous","_empty","_lowaqual","_notaligned") |
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172 |
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173 exclude <- match(c(exclude_HTSeq, exclude_DEXSeq),rownames(read_counts)) |
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174 exclude <- exclude[is.na(exclude)==0] |
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175 if(length(exclude) != 0) { |
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176 read_counts <- read_counts[-exclude,] |
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177 } |
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178 |
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179 |
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180 ## sorting expression matrix with the order of the read_counts |
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181 ##order <- match(colnames(read_counts) , rownames(design_matrix)) |
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182 ##read_counts_ordered <- read_counts[,order2] |
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183 |
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184 empty_samples <- apply(read_counts,2,function(x) sum(x) == 0) |
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185 if(sum(empty_samples) > 0) { |
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186 write(paste("There are ",sum(empty_samples)," empty samples found:",sep=""),stderr()) |
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187 write(colnames(read_counts)[empty_samples],stderr()) |
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188 } else { |
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189 |
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190 dge <- DGEList(counts=read_counts,genes=rownames(read_counts)) |
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191 |
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192 formula <- paste(c("~0",make.names(colnames(design_matrix))),collapse = " + ") |
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193 design_matrix_tmp <- design_matrix |
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194 colnames(design_matrix_tmp) <- make.names(colnames(design_matrix_tmp)) |
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195 design <- model.matrix(as.formula(formula),design_matrix_tmp) |
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196 rm(design_matrix_tmp) |
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197 |
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198 # Filter prefixes |
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199 prefixes = colnames(design_matrix)[attr(design,"assign")] |
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200 avoid = nchar(prefixes) == nchar(colnames(design)) |
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201 replacements = substr(colnames(design),nchar(prefixes)+1,nchar(colnames(design))) |
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202 replacements[avoid] = colnames(design)[avoid] |
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203 colnames(design) = replacements |
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204 |
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205 # Do normalization |
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206 write("Calculating normalization factors...",stdout()) |
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207 dge <- calcNormFactors(dge) |
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208 write("Estimating common dispersion...",stdout()) |
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209 dge <- estimateGLMCommonDisp(dge,design) |
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210 write("Estimating trended dispersion...",stdout()) |
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211 dge <- estimateGLMTrendedDisp(dge,design) |
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212 write("Estimating tagwise dispersion...",stdout()) |
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213 dge <- estimateGLMTagwiseDisp(dge,design) |
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214 |
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215 |
2
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216 # hierarchical clustering makes use of the distance of the MDS |
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217 if(output_MDSplot_logFC != "/dev/null" || output_hierarchical_clustering_plot != "/dev/null") { |
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218 write("Calculating MDS plot (logFC method)",stdout()) |
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219 mds_distance_logFC <- plotMDS.DGEList(dge,top=500,labels=rep("",nrow(dge\$samples)))# Get coordinates of unflexible plot |
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220 dev.off()# Kill it |
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221 |
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222 if(output_MDSplot_logFC != "/dev/null") { |
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223 write("Creating MDS plot (logFC method)",stdout()) |
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224 if(output_format_images == "pdf") { |
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225 pdf(output_MDSplot_logFC,height=14,width=14) |
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226 } else if(output_format_images == "svg") { |
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227 svg(output_MDSplot_logFC,height=14,width=14) |
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228 } else { |
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229 ## png(output_MDSplot_logFC) |
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230 ## png does not work out of the box in the Galaxy Toolshed Version of R due to its compile settings: https://biostar.usegalaxy.org/p/9170/ |
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231 |
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232 bitmap(output_MDSplot_logFC,type="png16m",height=7*3,width=7*3) |
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233 } |
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234 |
2
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235 diff_x <- abs(max(mds_distance_logFC\$x)-min(mds_distance_logFC\$x)) |
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236 diff_y <-(max(mds_distance_logFC\$y)-min(mds_distance_logFC\$y)) |
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237 plot(c(min(mds_distance_logFC\$x),max(mds_distance_logFC\$x) + 0.45 * diff_x), c(min(mds_distance_logFC\$y) - 0.05 * diff_y,max(mds_distance_logFC\$y) + 0.05 * diff_y), main="edgeR logFC-MDS Plot on top 500 genes",type="n", xlab="Leading logFC dim 1", ylab="Leading logFC dim 2") |
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238 points(mds_distance_logFC\$x,mds_distance_logFC\$y,pch=20) |
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239 text(mds_distance_logFC\$x, mds_distance_logFC\$y,rownames(dge\$samples),cex=1.25,col="gray",pos=4) |
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240 rm(diff_x,diff_y) |
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241 |
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242 dev.off() |
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243 } |
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244 } |
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245 |
2
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246 |
1
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247 if(output_MDSplot_bcv != "/dev/null") { |
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248 write("Creating MDS plot (bcv method)",stdout()) |
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249 |
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250 ## 1. First create a virtual plot to obtain the desired coordinates |
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251 pdf("bcvmds.pdf") |
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252 mds_distance_BCV <- plotMDS.DGEList(dge,method="bcv",top=500,labels=rep("",nrow(dge\$samples))) |
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253 dev.off()# Kill it |
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254 |
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255 ## 2. Re-plot the coordinates in a new figure with the size and settings. |
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256 if(output_format_images == "pdf") { |
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257 pdf(output_MDSplot_bcv,height=14,width=14) |
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258 } else if(output_format_images == "svg") { |
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259 svg(output_MDSplot_bcv,height=14,width=14) |
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260 } else { |
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261 ## png(output_MDSplot_bcv) |
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262 ## png does not work out of the box in the Galaxy Toolshed Version of R due to its compile settings: https://biostar.usegalaxy.org/p/9170/ |
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263 |
2
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264 bitmap(output_MDSplot_bcv,type="png16m",height=7*3,width=7*3) |
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265 } |
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266 |
2
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267 diff_x <- abs(max(mds_distance_BCV\$x)-min(mds_distance_BCV\$x)) |
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268 diff_y <- (max(mds_distance_BCV\$y)-min(mds_distance_BCV\$y)) |
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269 plot(c(min(mds_distance_BCV\$x),max(mds_distance_BCV\$x) + 0.45 * diff_x), c(min(mds_distance_BCV\$y) - 0.05 * diff_y,max(mds_distance_BCV\$y) + 0.05 * diff_y), main="edgeR BCV-MDS Plot",type="n", xlab="Leading BCV dim 1", ylab="Leading BCV dim 2") |
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270 points(mds_distance_BCV\$x,mds_distance_BCV\$y,pch=20) |
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271 text(mds_distance_BCV\$x, mds_distance_BCV\$y,rownames(dge\$samples),cex=1.25,col="gray",pos=4) |
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272 rm(diff_x,diff_y) |
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273 |
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274 dev.off() |
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275 } |
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276 |
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277 |
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278 if(output_BCVplot != "/dev/null") { |
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279 write("Creating Biological coefficient of variation plot",stdout()) |
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280 |
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281 if(output_format_images == "pdf") { |
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282 pdf(output_BCVplot) |
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283 } else if(output_format_images == "svg") { |
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284 svg(output_BCVplot) |
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285 } else { |
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286 ## png(output_BCVplot) |
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287 ## png does not work out of the box in the Galaxy Toolshed Version of R due to its compile settings: https://biostar.usegalaxy.org/p/9170/ |
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288 |
2
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289 bitmap(output_BCVplot,type="png16m",width=10.5*3,height=7*3) |
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290 } |
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291 |
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292 plotBCV(dge, cex=0.4, main="edgeR: Biological coefficient of variation (BCV) vs abundance") |
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293 dev.off() |
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294 } |
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295 |
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296 |
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297 write("Fitting GLM...",stdout()) |
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298 fit <- glmFit(dge,design) |
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299 |
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300 write(paste("Performing likelihood ratio test: ",contrast,sep=""),stdout()) |
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301 cont <- c(contrast) |
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302 cont <- makeContrasts(contrasts=cont, levels=design) |
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303 |
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304 lrt <- glmLRT(fit, contrast=cont[,1]) |
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305 write(paste("Exporting DGE results to file...",output_count_edgeR,sep=""),stdout()) |
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306 write.table(file=output_count_edgeR,topTags(lrt,n=nrow(read_counts))\$table,sep="\t",row.names=TRUE,col.names=NA) |
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307 write.table(file=output_cpm,cpm(dge,normalized.lib.sizes=TRUE),sep="\t",row.names=TRUE,col.names=NA) |
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308 |
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309 ## todo EXPORT FPKM |
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310 write.table(file=output_raw_counts,dge\$counts,sep="\t",row.names=TRUE,col.names=NA) |
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311 |
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312 if(output_MAplot != "/dev/null" || output_PValue_distribution_plot != "/dev/null") { |
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313 etable <- topTags(lrt, n=nrow(dge))\$table |
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314 etable <- etable[order(etable\$FDR), ] |
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315 |
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316 if(output_MAplot != "/dev/null") { |
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317 write("Creating MA plot...",stdout()) |
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318 |
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319 if(output_format_images == "pdf") { |
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320 pdf(output_MAplot) |
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321 } else if(output_format_images == "svg") { |
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322 svg(output_MAplot) |
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323 } else { |
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324 ## png(output_MAplot) |
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325 ## png does not work out of the box in the Galaxy Toolshed Version of R due to its compile settings: https://biostar.usegalaxy.org/p/9170/ |
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326 |
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327 bitmap(output_MAplot,type="png16m",width=10.5*3,height=7*3) |
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328 } |
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329 |
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330 with(etable, plot(logCPM, logFC, pch=20, main="edgeR: Fold change vs abundance")) |
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331 with(subset(etable, FDR < fdr), points(logCPM, logFC, pch=20, col="red")) |
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332 abline(h=c(-1,1), col="blue") |
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333 dev.off() |
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334 } |
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335 |
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336 if(output_PValue_distribution_plot != "/dev/null") { |
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337 write("Creating P-value distribution plot...",stdout()) |
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338 |
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339 if(output_format_images == "pdf") { |
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340 pdf(output_PValue_distribution_plot,width=14,height=14) |
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341 } else if(output_format_images == "svg") { |
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342 svg(output_PValue_distribution_plot,width=14,height=14) |
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343 } else { |
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344 ## png(output_PValue_distribution_plot) |
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345 ## png does not work out of the box in the Galaxy Toolshed Version of R due to its compile settings: https://biostar.usegalaxy.org/p/9170/ |
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346 |
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347 bitmap(output_PValue_distribution_plot,type="png16m",width=7*3,height=7*3) |
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348 } |
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349 |
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350 expressed_genes <- subset(etable, PValue < 0.99) |
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351 h <- hist(expressed_genes\$PValue,breaks=nrow(expressed_genes)/15,main="Binned P-Values (< 0.99)") |
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352 center <- sum(h\$counts) / length(h\$counts) |
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353 lines(c(0,1),c(center,center),lty=2,col="red",lwd=2) |
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354 k <- ksmooth(h\$mid, h\$counts) |
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355 lines(k\$x,k\$y,col="red",lwd=2) |
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356 rmsd <- (h\$counts) - center |
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357 rmsd <- rmsd^2 |
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358 rmsd <- sum(rmsd) |
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359 rmsd <- sqrt(rmsd) |
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360 text(0,max(h\$counts),paste("e=",round(rmsd,2),sep=""),pos=4,col="blue") |
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361 ## change e into epsilon somehow |
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362 dev.off() |
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363 } |
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364 } |
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365 |
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366 if(output_heatmap_plot != "/dev/null") { |
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367 |
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368 if(output_format_images == "pdf") { |
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369 pdf(output_heatmap_plot,width=10.5) |
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370 } else if(output_format_images == "svg") { |
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371 svg(output_heatmap_plot,width=10.5) |
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372 } else { |
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373 ## png(output_heatmap_plot) |
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374 ## png does not work out of the box in the Galaxy Toolshed Version of R due to its compile settings: https://biostar.usegalaxy.org/p/9170/ |
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375 |
2
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376 bitmap(output_heatmap_plot,type="png16m",width=10.5*3,height=7*3) |
1
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377 } |
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378 |
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379 etable2 <- topTags(lrt, n=100)\$table |
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380 order <- rownames(etable2) |
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381 cpm_sub <- cpm(dge,normalized.lib.sizes=TRUE,log=TRUE)[as.numeric(order),] |
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382 heatmap(t(cpm_sub)) |
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383 dev.off() |
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384 } |
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385 |
2
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386 if(output_hierarchical_clustering_plot != "/dev/null") { |
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387 if(output_hierarchical_clustering_plot == "pdf") { |
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388 pdf(output_hierarchical_clustering_plot,width=10.5) |
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389 } else if(output_hierarchical_clustering_plot == "svg") { |
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390 svg(output_hierarchical_clustering_plot,width=10.5) |
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391 } else { |
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392 ## png(output_hierarchical_clustering_plot) |
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393 ## png does not work out of the box in the Galaxy Toolshed Version of R due to its compile settings: https://biostar.usegalaxy.org/p/9170/ |
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394 |
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395 bitmap(output_hierarchical_clustering_plot,type="png16m",width=10.5*3,height=7*3) |
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396 } |
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397 |
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398 mds_distance = as.dist(mds_distance_logFC\$distance.matrix) |
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399 clustering = hclust(mds_distance) |
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400 plot(clustering,main=paste("Cluster Dendogram on the ",mds_distance_logFC\$top," TopTags",sep="",sub="\ncomplete linkage on logFC MDS distance")) |
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401 |
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402 dev.off() |
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403 } |
1
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404 |
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405 if(output_RData_obj != "/dev/null") { |
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406 save.image(output_RData_obj) |
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407 } |
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408 |
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409 write("Done!",stdout()) |
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410 } |
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411 ]]> |
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412 </configfile> |
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413 </configfiles> |
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414 |
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415 <inputs> |
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416 <param name="expression_matrix" type="data" format="tabular" label="Expression (read count) matrix" /> |
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417 <param name="design_matrix" type="data" format="tabular" label="Design matrix" help="Ensure your samplenames are identical to those in the expression matrix. Preferentially, create the contrast matrix using 'edgeR: Design- from Expression matrix'." /> |
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418 |
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419 <param name="contrast" type="text" label="Contrast (biological question)" help="e.g. 'tumor-normal' or '(G1+G2)/2-G3' using the factors chosen in the design matrix. Read the 'makeContrasts' manual from Limma package for more info: http://www.bioconductor.org/packages/release/bioc/html/limma.html and http://www.bioconductor.org/packages/release/bioc/vignettes/limma/inst/doc/usersguide.pdf." /> |
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420 |
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421 <param name="fdr" type="float" min="0" max="1" value="0.05" label="False Discovery Rate (FDR)" /> |
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422 |
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423 <param name="outputs" type="select" label="Optional desired outputs" multiple="true" display="checkboxes"> |
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424 <option value="make_output_raw_counts">Raw counts table</option> |
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425 <option value="make_output_MDSplot_logFC">MDS-plot (logFC-method)</option> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
426 <option value="make_output_MDSplot_bcv">MDS-plot (BCV-method; much slower)</option> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
427 <option value="make_output_BCVplot">BCV-plot</option> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
428 <option value="make_output_MAplot">MA-plot</option> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
429 <option value="make_output_PValue_distribution_plot">P-Value distribution plot</option> |
2
ec951a5017f8
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit a6cf3ec153ca4a3846258a223d287ca125eea7be
yhoogstrate
parents:
1
diff
changeset
|
430 <option value="make_output_hierarchical_clustering_plot">Hierarchical custering</option> |
1
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
431 <option value="make_output_heatmap_plot">Heatmap</option> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
432 <option value="make_output_RData_obj">R Data object</option> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
433 </param> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
434 |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
435 <param name="output_format_images" type="select" label="Output format of images" display="radio"> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
436 <option value="png">Portable network graphics (.png)</option> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
437 <option value="pdf">Portable document format (.pdf)</option> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
438 <option value="svg">Scalable vector graphics (.svg)</option> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
439 </param> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
440 </inputs> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
441 |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
442 <outputs> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
443 <data format="tabular" name="output_count_edgeR" label="edgeR DGE on ${design_matrix.hid}: ${design_matrix.name} - differentially expressed genes" /> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
444 <data format="tabular" name="output_cpm" label="edgeR DGE on ${design_matrix.hid}: ${design_matrix.name} - CPM" /> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
445 |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
446 <data format="tabular" name="output_raw_counts" label="edgeR DGE on ${design_matrix.hid}: ${design_matrix.name} - raw counts"> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
447 <filter>outputs and ("make_output_raw_counts" in outputs)</filter> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
448 </data> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
449 |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
450 <data format="png" name="output_MDSplot_logFC" label="edgeR DGE on ${design_matrix.hid}: ${design_matrix.name} - MDS-plot (logFC method)"> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
451 <filter>outputs and ("make_output_MDSplot_logFC" in outputs)</filter> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
452 |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
453 <change_format> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
454 <when input="output_format_images" value="png" format="png" /> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
455 <when input="output_format_images" value="pdf" format="pdf" /> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
456 <when input="output_format_images" value="svg" format="svg" /> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
457 </change_format> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
458 </data> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
459 |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
460 <data format="png" name="output_MDSplot_bcv" label="edgeR DGE on ${design_matrix.hid}: ${design_matrix.name} - MDS-plot (bcv method)"> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
461 <filter>outputs and ("make_output_MDSplot_bcv" in outputs)</filter> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
462 |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
463 <change_format> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
464 <when input="output_format_images" value="png" format="png" /> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
465 <when input="output_format_images" value="pdf" format="pdf" /> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
466 <when input="output_format_images" value="svg" format="svg" /> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
467 </change_format> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
468 </data> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
469 |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
470 <data format="png" name="output_BCVplot" label="edgeR DGE on ${design_matrix.hid}: ${design_matrix.name} - BCV-plot"> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
471 <filter>outputs and ("make_output_BCVplot" in outputs)</filter> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
472 |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
473 <change_format> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
474 <when input="output_format_images" value="png" format="png" /> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
475 <when input="output_format_images" value="pdf" format="pdf" /> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
476 <when input="output_format_images" value="svg" format="svg" /> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
477 </change_format> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
478 </data> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
479 |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
480 <data format="png" name="output_MAplot" label="edgeR DGE on ${design_matrix.hid}: ${design_matrix.name} - MA-plot"> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
481 <filter>outputs and ("make_output_MAplot" in outputs)</filter> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
482 |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
483 <change_format> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
484 <when input="output_format_images" value="png" format="png" /> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
485 <when input="output_format_images" value="pdf" format="pdf" /> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
486 <when input="output_format_images" value="svg" format="svg" /> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
487 </change_format> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
488 </data> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
489 |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
490 <data format="png" name="output_PValue_distribution_plot" label="edgeR DGE on ${design_matrix.hid}: ${design_matrix.name} - P-Value distribution"> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
491 <filter>outputs and ("make_output_PValue_distribution_plot" in outputs)</filter> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
492 |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
493 <change_format> |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
yhoogstrate
parents:
diff
changeset
|
494 <when input="output_format_images" value="png" format="png" /> |
a4a4c88783ea
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495 <when input="output_format_images" value="pdf" format="pdf" /> |
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|
496 <when input="output_format_images" value="svg" format="svg" /> |
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|
497 </change_format> |
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498 </data> |
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499 |
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500 <data format="png" name="output_hierarchical_clustering_plot" label="edgeR DGE on ${design_matrix.hid}: ${design_matrix.name} - Hierarchical custering"> |
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501 <filter>outputs and ("make_output_hierarchical_clustering_plot" in outputs)</filter> |
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502 |
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503 <change_format> |
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504 <when input="output_format_images" value="png" format="png" /> |
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505 <when input="output_format_images" value="pdf" format="pdf" /> |
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506 <when input="output_format_images" value="svg" format="svg" /> |
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507 </change_format> |
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508 </data> |
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509 |
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510 <data format="png" name="output_heatmap_plot" label="edgeR DGE on ${design_matrix.hid}: ${design_matrix.name} - Heatmap"> |
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511 <filter>outputs and ("make_output_heatmap_plot" in outputs)</filter> |
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512 |
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513 <change_format> |
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514 <when input="output_format_images" value="png" format="png" /> |
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515 <when input="output_format_images" value="pdf" format="pdf" /> |
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516 <when input="output_format_images" value="svg" format="svg" /> |
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517 </change_format> |
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518 </data> |
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519 |
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520 <data format="RData" name="output_RData_obj" label="edgeR DGE on ${design_matrix.hid}: ${design_matrix.name} - R data object"> |
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521 <filter>outputs and ("make_output_RData_obj" in outputs)</filter> |
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522 </data> |
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523 |
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524 <data format="txt" name="output_R" label="edgeR DGE on ${design_matrix.hid}: ${design_matrix.name} - R output (debug)" > |
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525 <filter>outputs and ("make_output_R_stdout" in outputs)</filter> |
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526 </data> |
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527 </outputs> |
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528 |
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529 <tests> |
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530 <test> |
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531 <param name="expression_matrix" value="Differential_Gene_Expression/expression_matrix.tabular.txt" /> |
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532 <param name="design_matrix" value="Differential_Gene_Expression/design_matrix.tabular.txt" /> |
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533 |
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534 <param name="contrast" value="E-C"/> |
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535 |
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536 <param name="fdr" value="0.05" /> |
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537 |
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538 <param name="output_format_images" value="png" /> |
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539 |
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540 <output name="output_count_edgeR" file="Differential_Gene_Expression/differentially_expressed_genes.tabular.txt" /> |
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541 </test> |
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542 </tests> |
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543 |
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544 <help> |
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545 edgeR: Differential Gene(Expression) Analysis |
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546 ############################################# |
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547 |
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548 Overview |
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549 -------- |
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550 Differential expression analysis of RNA-seq and digital gene expression profiles with biological replication. Uses empirical Bayes estimation and exact tests based on the negative binomial distribution. Also useful for differential signal analysis with other types of genome-scale count data [1]. |
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551 |
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552 For every experiment, the algorithm requires a design matrix. This matrix describes which samples belong to which groups. |
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553 More details on this are given in the edgeR manual: http://www.bioconductor.org/packages/2.12/bioc/vignettes/edgeR/inst/doc/edgeRUsersGuide.pdf |
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554 and the limma manual. |
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555 |
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556 Because the creation of a design matrix can be complex and time consuming, especially if no GUI is used, this package comes with an alternative tool which can help you with it. |
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557 This tool is called *edgeR Design Matrix Creator*. |
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558 If the appropriate design matrix (with corresponding links to the files) is given, |
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559 the correct contrast ( http://en.wikipedia.org/wiki/Contrast_(statistics) ) has to be given. |
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560 |
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561 If you have for example two groups, with an equal weight, you would like to compare either |
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562 "g1-g2" or "normal-cancer". |
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563 |
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564 The test function makes use of a MCF7 dataset used in a study that indicates that a higher sequencing depth is not neccesairily more important than a higher amount of replaciates[2]. |
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565 |
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planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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566 Input |
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567 ----- |
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planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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568 Expression matrix |
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569 ^^^^^^^^^^^^^^^^^ |
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570 :: |
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571 |
a4a4c88783ea
planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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572 Geneid "\t" Sample-1 "\t" Sample-2 "\t" Sample-3 "\t" Sample-4 [...] "\n" |
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planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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573 SMURF "\t" 123 "\t" 21 "\t" 34545 "\t" 98 ... "\n" |
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574 BRCA1 "\t" 435 "\t" 6655 "\t" 45 "\t" 55 ... "\n" |
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planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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575 LINK33 "\t" 4 "\t" 645 "\t" 345 "\t" 1 ... "\n" |
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576 SNORD78 "\t" 498 "\t" 65 "\t" 98 "\t" 27 ... "\n" |
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planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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577 [...] |
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planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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578 |
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planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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579 *Note: Make sure the number of columns in the header is identical to the number of columns in the body.* |
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580 |
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planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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581 Design matrix |
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582 ^^^^^^^^^^^^^ |
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planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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583 :: |
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584 |
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planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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585 Sample "\t" Condition "\t" Ethnicity "\t" Patient "\t" Batch "\n" |
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586 Sample-1 "\t" Tumor "\t" European "\t" 1 "\t" 1 "\n" |
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587 Sample-2 "\t" Normal "\t" European "\t" 1 "\t" 1 "\n" |
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planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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588 Sample-3 "\t" Tumor "\t" European "\t" 2 "\t" 1 "\n" |
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planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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589 Sample-4 "\t" Normal "\t" European "\t" 2 "\t" 1 "\n" |
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590 Sample-5 "\t" Tumor "\t" African "\t" 3 "\t" 1 "\n" |
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planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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591 Sample-6 "\t" Normal "\t" African "\t" 3 "\t" 1 "\n" |
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592 Sample-7 "\t" Tumor "\t" African "\t" 4 "\t" 2 "\n" |
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593 Sample-8 "\t" Normal "\t" African "\t" 4 "\t" 2 "\n" |
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594 Sample-9 "\t" Tumor "\t" Asian "\t" 5 "\t" 2 "\n" |
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planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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595 Sample-10 "\t" Normal "\t" Asian "\t" 5 "\t" 2 "\n" |
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planemo upload for repository https://bitbucket.org/EMCbioinf/galaxy-tool-shed-tools/raw/master/edger_with_design_matrix commit 2700e500a4fb135a20ede7d52221a9d31f1aaa5e-dirty
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596 Sample-11 "\t" Tumor "\t" Asian "\t" 6 "\t" 2 "\n" |
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597 Sample-12 "\t" Normal "\t" Asian "\t" 6 "\t" 2 "\n" |
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598 |
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599 *Note: Avoid factor names that are (1) numerical, (2) contain mathematical symbols and preferebly only use letters.* |
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600 |
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601 Contrast |
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602 ^^^^^^^^ |
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603 The contrast represents the biological question. There can be many questions asked, e.g.: |
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604 |
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605 - Tumor-Normal |
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606 - African-European |
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607 - 0.5*(Control+Placebo) / Treated |
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608 |
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609 Installation |
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610 ------------ |
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611 |
2
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612 This tool requires no specific configuration. The following dependencies will installed automatically: |
1
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613 |
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614 - R |
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615 - limma |
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616 - edgeR |
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617 |
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618 License |
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619 ------- |
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620 - R |
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621 - GPL 2 & GPL 3 |
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622 - limma |
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623 - GPL (>=2) |
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624 - edgeR |
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625 - GPL (>=2) |
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626 |
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627 @CONTACT@ |
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628 </help> |
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629 |
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630 <expand macro="citations" /> |
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631 </tool> |