Mercurial > repos > iuc > scater_filter
annotate scater-plot-dist-scatter.R @ 1:b7ea9f09c02f draft
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
| author | iuc | 
|---|---|
| date | Tue, 03 Sep 2019 14:27:39 -0400 | 
| parents | e6ca62ac65c6 | 
| children | 7a365ec81b52 | 
| rev | line source | 
|---|---|
| 
0
 
e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
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 | 
1 #!/usr/bin/env Rscript | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
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 | 
2 | 
| 
 
e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
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 | 
3 # Plot the distribution of read counts and feature counts, side by side, then a scatter plot of read counts vs feature counts below | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
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 | 
4 | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
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 | 
5 # Load optparse we need to check inputs | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
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 | 
6 | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
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 | 
7 library(optparse) | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
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 | 
8 library(workflowscriptscommon) | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
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 | 
9 library(LoomExperiment) | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
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 | 
10 library(scater) | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
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11 library(ggpubr) | 
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1
 
b7ea9f09c02f
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
 
iuc 
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12 library(scales) | 
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0
 
e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
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13 | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
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14 # parse options | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
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15 | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
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16 option_list = list( | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
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diff
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 | 
17 make_option( | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
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18 c("-i", "--input-loom"), | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
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 | 
19 action = "store", | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
diff
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 | 
20 default = NA, | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
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diff
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 | 
21 type = 'character', | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
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 | 
22 help = "A SingleCellExperiment object file in Loom format." | 
| 
 
e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
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diff
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 | 
23 ), | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
diff
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 | 
24 make_option( | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
diff
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 | 
25 c("-o", "--output-plot-file"), | 
| 
 
e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
diff
changeset
 | 
26 action = "store", | 
| 
 
e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
diff
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27 default = NA, | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
diff
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 | 
28 type = 'character', | 
| 
 
e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
diff
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 | 
29 help = "Path of the PDF output file to save plot to." | 
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1
 
b7ea9f09c02f
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
 
iuc 
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30 ), | 
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b7ea9f09c02f
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
 
iuc 
parents: 
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31 make_option( | 
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b7ea9f09c02f
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
 
iuc 
parents: 
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32 c("-l", "--log-scale"), | 
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b7ea9f09c02f
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
 
iuc 
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33 action="store_true", | 
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b7ea9f09c02f
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
 
iuc 
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34 default=FALSE, | 
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b7ea9f09c02f
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
 
iuc 
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35 type = 'logical', | 
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b7ea9f09c02f
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
 
iuc 
parents: 
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36 help = "Plot on log scale (recommended for large datasets)." | 
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0
 
e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
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37 ) | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
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38 ) | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
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diff
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39 | 
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1
 
b7ea9f09c02f
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
 
iuc 
parents: 
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40 opt <- wsc_parse_args(option_list, mandatory = c('input_loom', 'output_plot_file', 'log_scale')) | 
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0
 
e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
diff
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41 | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
diff
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42 # Check parameter values | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
diff
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43 | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
diff
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44 if ( ! file.exists(opt$input_loom)){ | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
diff
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45 stop((paste('File', opt$input_loom, 'does not exist'))) | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
diff
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46 } | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
diff
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47 | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
diff
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48 # Input from Loom format | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
diff
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49 | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
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50 scle <- import(opt$input_loom, format='loom', type='SingleCellLoomExperiment') | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
diff
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51 | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
diff
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52 #do the scatter plot of reads vs genes | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
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53 total_counts <- scle$total_counts | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
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54 total_features <- scle$total_features_by_counts | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
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55 count_feats <- cbind(total_counts, total_features) | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
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56 cf_dm <- as.data.frame(count_feats) | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
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57 | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
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58 # Calculate binwidths for reads and features plots. Use 20 bins | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
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59 read_bins <- max(total_counts / 1e6) / 20 | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
diff
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60 feat_bins <- max(total_features) / 20 | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
parents:  
diff
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61 | 
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1
 
b7ea9f09c02f
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
 
iuc 
parents: 
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62 # Make the plots | 
| 
0
 
e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
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63 plot <- ggplot(cf_dm, aes(x=total_counts / 1e6, y=total_features)) + geom_point(shape=1) + geom_smooth() + xlab("Read count (millions)") + | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
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64 ylab("Feature count") + ggtitle("Scatterplot of reads vs features") | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
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65 plot1 <- qplot(total_counts / 1e6, geom="histogram", binwidth = read_bins, ylab="Number of cells", xlab = "Read counts (millions)", fill=I("darkseagreen3")) + ggtitle("Read counts per cell") | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
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66 plot2 <- qplot(total_features, geom="histogram", binwidth = feat_bins, ylab="Number of cells", xlab = "Feature counts", fill=I("darkseagreen3")) + ggtitle("Feature counts per cell") | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
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67 plot3 <- plotColData(scle, y="pct_counts_MT", x="total_features_by_counts") + ggtitle("% MT genes") + geom_point(shape=1) + theme(text = element_text(size=15)) + theme(plot.title = element_text(size=15)) | 
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e6ca62ac65c6
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 5fdcafccb6c645d301db040dfeed693d7b6b4278
 
iuc 
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68 | 
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1
 
b7ea9f09c02f
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
 
iuc 
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69 if (! opt$log_scale){ | 
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b7ea9f09c02f
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
 
iuc 
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70 final_plot <- ggarrange(plot1, plot2, plot, plot3, ncol=2, nrow=2) | 
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b7ea9f09c02f
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
 
iuc 
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71 ggsave(opt$output_plot_file, final_plot, device="pdf") | 
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b7ea9f09c02f
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
 
iuc 
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72 } else { | 
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b7ea9f09c02f
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
 
iuc 
parents: 
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73 plot_log_both <- plot + scale_x_continuous(trans = 'log10') + scale_y_continuous(trans = 'log10') | 
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b7ea9f09c02f
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
 
iuc 
parents: 
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74 plot1_log <- plot1 + scale_y_continuous(trans = 'log10') | 
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b7ea9f09c02f
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
 
iuc 
parents: 
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75 plot2_log <- plot2 + scale_y_continuous(trans = 'log10') | 
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b7ea9f09c02f
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
 
iuc 
parents: 
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76 plot3_log <- plot3 + scale_y_log10(labels=number) | 
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b7ea9f09c02f
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
 
iuc 
parents: 
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77 final_plot_log <- ggarrange(plot1_log, plot2_log, plot_log_both, plot3_log, ncol=2, nrow=2) | 
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b7ea9f09c02f
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
 
iuc 
parents: 
0 
diff
changeset
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78 ggsave(opt$output_plot_file, final_plot_log, device="pdf") | 
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b7ea9f09c02f
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
 
iuc 
parents: 
0 
diff
changeset
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79 } | 
