Mercurial > repos > artbio > repenrich
annotate edgeR_repenrich.R @ 15:2e3d976e7d5d draft default tip
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 03183e29f807ec33548016a7c4144f52720b7b9e
author | artbio |
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date | Sun, 21 Apr 2024 09:44:51 +0000 |
parents | 530626b0757c |
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f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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1 # setup R error handling to go to stderr |
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6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
artbio
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2 options(show.error.messages = FALSE, error = function() { |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
artbio
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3 cat(geterrmessage(), file = stderr()) |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
artbio
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4 q("no", 1, FALSE) |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
artbio
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5 }) |
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f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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6 |
f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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7 # To not crash galaxy with an UTF8 error with not-US LC settings. |
f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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8 loc <- Sys.setlocale("LC_MESSAGES", "en_US.UTF-8") |
f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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9 |
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6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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10 # load libraries |
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f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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11 library("getopt") |
f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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12 library("tools") |
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6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
artbio
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13 library("rjson") |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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14 suppressPackageStartupMessages({ |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
artbio
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15 library("edgeR") |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
artbio
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16 library("limma") |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
artbio
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17 }) |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
artbio
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18 |
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f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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19 options(stringAsFactors = FALSE, useFancyQuotes = FALSE) |
f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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20 |
f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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21 # get options, using the spec as defined by the enclosed list. |
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6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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22 spec <- matrix( |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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23 c( |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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24 "quiet", "q", 0, "logical", |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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25 "outfile", "o", 1, "character", |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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26 "countsfile", "n", 1, "character", |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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27 "factorName", "N", 1, "character", |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
artbio
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28 "levelNameA", "A", 1, "character", |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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29 "levelNameB", "B", 1, "character", |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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30 "levelAfiles", "a", 1, "character", |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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31 "levelBfiles", "b", 1, "character", |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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32 "plots", "p", 1, "character" |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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33 ), |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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34 byrow = TRUE, ncol = 4 |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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35 ) |
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f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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36 opt <- getopt(spec) |
f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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37 |
f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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38 # build levels A and B file lists |
f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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39 filesA <- fromJSON(opt$levelAfiles, method = "C", unexpected.escape = "error") |
f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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40 filesB <- fromJSON(opt$levelBfiles, method = "C", unexpected.escape = "error") |
f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
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41 listA <- list() |
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planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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42 indice <- 0 |
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f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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43 listA[["level"]] <- opt$levelNameA |
f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
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44 for (file in filesA) { |
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6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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45 indice <- indice + 1 |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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46 listA[[paste0(opt$levelNameA, "_", indice)]] <- read.delim(file, header = FALSE) |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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47 } |
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planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
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48 listB <- list() |
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planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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49 indice <- 0 |
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f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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50 listB[["level"]] <- opt$levelNameB |
f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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51 for (file in filesB) { |
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planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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52 indice <- indice + 1 |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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53 listB[[paste0(opt$levelNameB, "_", indice)]] <- read.delim(file, header = FALSE) |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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54 } |
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f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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55 |
f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
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56 # build a counts table |
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planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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57 counts <- data.frame(row.names = listA[[2]][, 1]) |
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f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
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58 for (element in names(listA[-1])) { |
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planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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59 counts <- cbind(counts, listA[[element]][, 4]) |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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60 } |
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f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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61 for (element in names(listB[-1])) { |
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6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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62 counts <- cbind(counts, listB[[element]][, 4]) |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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63 } |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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64 colnames(counts) <- c(names(listA[-1]), names(listB[-1])) |
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planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit df6b9491ad06e8a85e67c663b68db3cce3eb0115
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65 sizes <- colSums(counts) |
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f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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66 |
f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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67 # build a meta data object |
f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
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68 meta <- data.frame( |
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planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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69 row.names = colnames(counts), |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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70 condition = c(rep(opt$levelNameA, length(filesA)), rep(opt$levelNameB, length(filesB))), |
6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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71 libsize = sizes |
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f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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72 ) |
f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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73 |
f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
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74 |
f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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75 # Define the library size and conditions for the GLM |
f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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76 libsize <- meta$libsize |
f6f0f1e5e940
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/master/tools/repenrich commit 61e203df0be5ed877ff92b917c7cde6eeeab8310
artbio
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77 condition <- factor(meta$condition) |
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6bba3e33c2e7
planemo upload for repository https://github.com/ARTbio/tools-artbio/tree/main/tools/repenrich commit 11df9f0b68d35d3a9424a17e4cefee6cfb9d4c19
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78 design <- model.matrix(~ 0 + condition) |
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79 colnames(design) <- levels(condition) |
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80 |
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81 # Build a DGE object for the GLM |
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82 y <- DGEList(counts = counts, lib.size = libsize) |
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83 |
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84 # Normalize the data |
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85 y <- calcNormFactors(y) |
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86 |
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87 # Estimate the variance |
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88 y <- estimateGLMCommonDisp(y, design) |
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89 y <- estimateGLMTrendedDisp(y, design) |
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90 y <- estimateGLMTagwiseDisp(y, design) |
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91 |
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92 # Builds and outputs an object to contain the normalized read abundance in counts per million of reads |
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93 cpm <- cpm(y, log = FALSE, lib.size = libsize) |
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94 cpm <- as.data.frame(cpm) |
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95 colnames(cpm) <- colnames(counts) |
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96 if (!is.null(opt$countsfile)) { |
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97 normalizedAbundance <- data.frame(Tag = rownames(cpm)) |
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98 normalizedAbundance <- cbind(normalizedAbundance, cpm) |
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99 write.table(normalizedAbundance, file = opt$countsfile, sep = "\t", col.names = TRUE, row.names = FALSE, quote = FALSE) |
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100 } |
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101 |
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102 # Conduct fitting of the GLM |
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103 yfit <- glmFit(y, design) |
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104 |
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105 # Initialize result matrices to contain the results of the GLM |
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106 results <- matrix(nrow = dim(counts)[1], ncol = 0) |
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107 logfc <- matrix(nrow = dim(counts)[1], ncol = 0) |
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108 |
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109 # Make the comparisons for the GLM |
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110 my.contrasts <- makeContrasts( |
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111 paste0(opt$levelNameA, "_", opt$levelNameB, " = ", opt$levelNameA, " - ", opt$levelNameB), |
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112 levels = design |
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113 ) |
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114 |
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115 # Define the contrasts used in the comparisons |
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116 allcontrasts <- paste0(opt$levelNameA, " vs ", opt$levelNameB) |
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117 |
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118 # Conduct a for loop that will do the fitting of the GLM for each comparison |
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119 # Put the results into the results objects |
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120 lrt <- glmLRT(yfit, contrast = my.contrasts[, 1]) |
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121 res <- topTags(lrt, n = dim(c)[1], sort.by = "none")$table |
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122 results <- cbind(results, res[, c(1, 5)]) |
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123 logfc <- cbind(logfc, res[c(1)]) |
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124 |
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125 # Add the repeat types back into the results. |
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126 # We should still have the same order as the input data |
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127 results$class <- listA[[2]][, 2] |
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128 results$type <- listA[[2]][, 3] |
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129 # Sort the results table by the FDR |
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130 results <- results[with(results, order(FDR)), ] |
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131 |
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132 # Plot Fold Changes for repeat classes and types |
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133 |
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134 # open the device and plots |
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135 if (!is.null(opt$plots)) { |
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136 pdf(opt$plots) |
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137 plotMDS(y, main = "Multidimensional Scaling Plot Of Distances Between Samples") |
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138 plotBCV(y, xlab = "Gene abundance (Average log CPM)", main = "Biological Coefficient of Variation Plot") |
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139 logFC <- results[, "logFC"] |
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140 # Plot the repeat classes |
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141 classes <- with(results, reorder(class, -logFC, median)) |
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142 classes |
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143 par(mar = c(6, 10, 4, 1)) |
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144 boxplot(logFC ~ classes, |
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145 data = results, outline = FALSE, horizontal = TRUE, |
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146 las = 2, xlab = "log2(Fold Change)", ylab = "", cex.axis = 0.7, main = paste0(allcontrasts, ", by Class") |
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147 ) |
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148 abline(v = 0) |
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149 # Plot the repeat types |
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150 types <- with(results, reorder(type, -logFC, median)) |
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151 boxplot(logFC ~ types, |
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152 data = results, outline = FALSE, horizontal = TRUE, |
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153 las = 2, xlab = "log2(Fold Change)", ylab = "", cex.axis = 0.7, main = paste0(allcontrasts, ", by Type") |
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154 ) |
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155 abline(v = 0) |
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156 # volcano plot |
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157 TEdata <- cbind(rownames(results), as.data.frame(results), score = -log(results$FDR, 10)) |
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158 colnames(TEdata) <- c("Tag", "log2FC", "FDR", "Class", "Type", "score") |
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159 color <- ifelse(TEdata$FDR < 0.05, "red", "black") |
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160 s <- subset(TEdata, FDR < 0.01) |
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161 with(TEdata, plot(log2FC, score, pch = 20, col = color, main = "Volcano plot (all tag types)", ylab = "-log10(FDR)")) |
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162 text(s[, 2], s[, 6], labels = s[, 5], pos = seq(from = 1, to = 3), cex = 0.5) |
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163 } |
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164 |
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165 # close the plot device |
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166 if (!is.null(opt$plots)) { |
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167 cat("closing plot device\n") |
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168 dev.off() |
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169 } |
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170 |
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171 # Save the results |
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172 results <- cbind(TE_item = rownames(results), results) |
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173 colnames(results) <- c("TE_item", "log2FC", "FDR", "Class", "Type") |
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174 results$log2FC <- format(results$log2FC, digits = 5) |
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175 results$FDR <- format(results$FDR, digits = 5) |
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176 write.table(results, opt$outfile, quote = FALSE, sep = "\t", col.names = TRUE, row.names = FALSE) |