Mercurial > repos > mingchen0919 > rmarkdown_deseq2
annotate DESeq.Rmd @ 4:7f0281d26b81 draft
planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit a23e23222252167ef7c3338a4872e84706df8f83-dirty
author | mingchen0919 |
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date | Tue, 08 Aug 2017 14:33:57 -0400 |
parents | 312e9bcc02f1 |
children | 2f8ddef8d545 |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
mingchen0919
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1 --- |
7231d7e8d3ed
planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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2 title: 'DESeq2: Perform DESeq analysis' |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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3 output: |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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4 html_document: |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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5 number_sections: true |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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6 toc: true |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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7 theme: cosmo |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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8 highlight: tango |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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9 --- |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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10 |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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11 ```{r setup, include=FALSE, warning=FALSE, message=FALSE} |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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12 knitr::opts_chunk$set( |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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13 echo = ECHO |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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14 ) |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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15 |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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16 library(stringi) |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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17 library(DESeq2) |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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18 library(pheatmap) |
1
312e9bcc02f1
planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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19 # library(PoiClaClu) |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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20 library(RColorBrewer) |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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21 ``` |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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22 |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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23 # `DESeqDataSet` object |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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24 |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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25 ```{r} |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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26 count_files = strsplit(opt$count_files, ',')[[1]] |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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27 sample_table = read.table(opt$sample_table, header = TRUE) |
7231d7e8d3ed
planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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28 |
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29 ## copy count files into working directory |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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30 file_copy = file.copy(count_files, sample_table$fileName, overwrite = TRUE) |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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31 |
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32 ## DESeqDataSet object |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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33 dds = DESeqDataSetFromHTSeqCount(sampleTable = sample_table, |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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34 directory = './', |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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35 design = DESIGN_FORMULA) |
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planemo upload for repository https://github.com/statonlab/docker-GRReport/tree/master/my_tools/rmarkdown_deseq2 commit 9285c2b8ad41a486dde2a87600a6b8267841c8b5-dirty
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36 dds |
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37 ``` |
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38 |
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39 # Pre-filtering the dataset. |
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40 |
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41 We can remove the rows that have 0 or 1 count to reduce object size and increase the calculation speed. |
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42 |
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43 * Number of rows before pre-filtering |
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44 ```{r} |
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45 nrow(dds) |
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46 ``` |
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47 |
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48 * Number of rows after pre-filtering |
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49 ```{r} |
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50 dds = dds[rowSums(counts(dds)) > 1, ] |
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51 nrow(dds) |
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52 ``` |
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53 |
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54 # Peek at data {.tabset} |
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55 |
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56 ## Count Data |
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57 |
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58 ```{r} |
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59 datatable(head(counts(dds), 100), style="bootstrap", |
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60 class="table-condensed", options = list(dom = 'tp', scrollX = TRUE)) |
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61 ``` |
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62 |
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63 ## Sample Table |
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64 |
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65 ```{r} |
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66 datatable(sample_table, style="bootstrap", |
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67 class="table-condensed", options = list(dom = 'tp', scrollX = TRUE)) |
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68 ``` |
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69 |
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70 # Sample distance on variance stabilized data {.tabset} |
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71 |
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72 ## `rlog` Stabilizing transformation |
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73 |
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74 ```{r} |
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75 rld = rlog(dds, blind = FALSE) |
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76 datatable(head(assay(rld), 100), style="bootstrap", |
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77 class="table-condensed", options = list(dom = 'tp', scrollX = TRUE)) |
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78 ``` |
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79 |
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80 ## Sample distance |
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81 |
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82 ```{r} |
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83 sampleDists <- dist(t(assay(rld))) |
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84 sampleDists |
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85 ``` |
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86 |
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87 # Differential expression analysis |
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88 |
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89 ```{r} |
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90 dds <- DESeq(dds) |
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91 ``` |
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92 |
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93 ```{r} |
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94 rm("opt") |
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95 save(list=ls(all.names = TRUE), file='DESEQ_WORKSPACE') |
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96 ``` |
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97 |
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98 |