From 45799e7aebb62bde4a5543880f2ee107d9fb8496 Mon Sep 17 00:00:00 2001 From: Dominik <dominik.brilhaus@hhu.de> Date: Thu, 24 Mar 2022 20:56:42 +0100 Subject: [PATCH] workaround kallisto dependency --- README.md | 18 +- .../01_kallisto_results/DB_097/abundance.tsv | 0 .../01_kallisto_results/DB_097/run_info.json | 0 .../01_kallisto_results/DB_099/abundance.tsv | 0 .../01_kallisto_results/DB_099/run_info.json | 0 .../01_kallisto_results/DB_103/abundance.tsv | 0 .../01_kallisto_results/DB_103/run_info.json | 0 .../01_kallisto_results/DB_161/abundance.tsv | 0 .../01_kallisto_results/DB_161/run_info.json | 0 .../01_kallisto_results/DB_163/abundance.tsv | 0 .../01_kallisto_results/DB_163/run_info.json | 0 .../01_kallisto_results/DB_165/abundance.tsv | 0 .../01_kallisto_results/DB_165/run_info.json | 0 .../run1/03_kallisto_mappingStats.csv | 0 .../run1/03_kallisto_tpmMatrix.csv | 0 .../run1/04_sleuth_dge.csv | 0 .../run1/05_shinyPrep.RData | Bin runs/{ => kallisto_sleuth}/run1/isa.run.xlsx | Bin .../run2/03_kallisto_mappingStats.csv | 7 + .../run2/03_kallisto_tpmMatrix.csv | 31435 ++++++++++++++++ runs/kallisto_sleuth/run2/04_sleuth_dge.csv | 19714 ++++++++++ runs/kallisto_sleuth/run2/05_shinyPrep.RData | Bin 0 -> 1947166 bytes runs/run1/01_kallisto_index | 3 - workflows/.Rhistory | 131 + workflows/kallisto_sleuth/01_KallistoIndex.sh | 25 + workflows/kallisto_sleuth/01_KallistoQuant.sh | 24 +- .../kallisto_sleuth/02_InstallPackages.R | 10 +- .../kallisto_sleuth/03_KallistoCollect.R | 9 +- workflows/kallisto_sleuth_wrapper.sh | 31 +- workflows/plot_shinyApp.Rmd | 2 +- 30 files changed, 51372 insertions(+), 37 deletions(-) rename runs/{ => kallisto_sleuth}/run1/01_kallisto_results/DB_097/abundance.tsv (100%) rename runs/{ => kallisto_sleuth}/run1/01_kallisto_results/DB_097/run_info.json (100%) rename runs/{ => kallisto_sleuth}/run1/01_kallisto_results/DB_099/abundance.tsv (100%) rename runs/{ => kallisto_sleuth}/run1/01_kallisto_results/DB_099/run_info.json (100%) rename runs/{ => kallisto_sleuth}/run1/01_kallisto_results/DB_103/abundance.tsv (100%) rename runs/{ => kallisto_sleuth}/run1/01_kallisto_results/DB_103/run_info.json (100%) rename runs/{ => kallisto_sleuth}/run1/01_kallisto_results/DB_161/abundance.tsv (100%) rename runs/{ => kallisto_sleuth}/run1/01_kallisto_results/DB_161/run_info.json (100%) rename runs/{ => kallisto_sleuth}/run1/01_kallisto_results/DB_163/abundance.tsv (100%) rename runs/{ => kallisto_sleuth}/run1/01_kallisto_results/DB_163/run_info.json (100%) rename runs/{ => kallisto_sleuth}/run1/01_kallisto_results/DB_165/abundance.tsv (100%) rename runs/{ => kallisto_sleuth}/run1/01_kallisto_results/DB_165/run_info.json (100%) rename runs/{ => kallisto_sleuth}/run1/03_kallisto_mappingStats.csv (100%) rename runs/{ => kallisto_sleuth}/run1/03_kallisto_tpmMatrix.csv (100%) rename runs/{ => kallisto_sleuth}/run1/04_sleuth_dge.csv (100%) rename runs/{ => kallisto_sleuth}/run1/05_shinyPrep.RData (100%) rename runs/{ => kallisto_sleuth}/run1/isa.run.xlsx (100%) create mode 100644 runs/kallisto_sleuth/run2/03_kallisto_mappingStats.csv create mode 100644 runs/kallisto_sleuth/run2/03_kallisto_tpmMatrix.csv create mode 100644 runs/kallisto_sleuth/run2/04_sleuth_dge.csv create mode 100644 runs/kallisto_sleuth/run2/05_shinyPrep.RData delete mode 100644 runs/run1/01_kallisto_index create mode 100644 workflows/.Rhistory create mode 100755 workflows/kallisto_sleuth/01_KallistoIndex.sh mode change 100644 => 100755 workflows/kallisto_sleuth/01_KallistoQuant.sh diff --git a/README.md b/README.md index a92020d..3fd845d 100644 --- a/README.md +++ b/README.md @@ -103,4 +103,20 @@ echo "~/03DataPLANT_gitlab/samplearc_rnaseq/" > workflows/_arc_local_wd # replace -# runs/ by runs/'$run_name'/ \ No newline at end of file +# runs/ by runs/'$run_name'/ + + +# PKG_PATH="http://cdn.NetBSD.org/pub/pkgsrc/packages/NetBSD/$(uname -p)/$(uname -r|cut -f '1 2' -d.)/All/" +# export PKG_PATH +# pkg_add pkgin + + + +# git clone https://github.com/pachterlab/kallisto.git +# cd kallisto +# mkdir build +# cd build +# cmake .. -DUSE_HDF5=ON + +-- Build files have been written to: /Users/dominikbrilhaus/kallisto/build + diff --git a/runs/run1/01_kallisto_results/DB_097/abundance.tsv b/runs/kallisto_sleuth/run1/01_kallisto_results/DB_097/abundance.tsv similarity index 100% rename from runs/run1/01_kallisto_results/DB_097/abundance.tsv rename to runs/kallisto_sleuth/run1/01_kallisto_results/DB_097/abundance.tsv diff --git a/runs/run1/01_kallisto_results/DB_097/run_info.json b/runs/kallisto_sleuth/run1/01_kallisto_results/DB_097/run_info.json similarity index 100% rename from runs/run1/01_kallisto_results/DB_097/run_info.json rename to runs/kallisto_sleuth/run1/01_kallisto_results/DB_097/run_info.json diff --git a/runs/run1/01_kallisto_results/DB_099/abundance.tsv b/runs/kallisto_sleuth/run1/01_kallisto_results/DB_099/abundance.tsv similarity index 100% rename from runs/run1/01_kallisto_results/DB_099/abundance.tsv rename to runs/kallisto_sleuth/run1/01_kallisto_results/DB_099/abundance.tsv diff --git a/runs/run1/01_kallisto_results/DB_099/run_info.json b/runs/kallisto_sleuth/run1/01_kallisto_results/DB_099/run_info.json similarity index 100% rename from runs/run1/01_kallisto_results/DB_099/run_info.json rename to runs/kallisto_sleuth/run1/01_kallisto_results/DB_099/run_info.json diff --git a/runs/run1/01_kallisto_results/DB_103/abundance.tsv b/runs/kallisto_sleuth/run1/01_kallisto_results/DB_103/abundance.tsv similarity index 100% rename from runs/run1/01_kallisto_results/DB_103/abundance.tsv rename to runs/kallisto_sleuth/run1/01_kallisto_results/DB_103/abundance.tsv diff --git a/runs/run1/01_kallisto_results/DB_103/run_info.json b/runs/kallisto_sleuth/run1/01_kallisto_results/DB_103/run_info.json similarity index 100% rename from runs/run1/01_kallisto_results/DB_103/run_info.json rename to runs/kallisto_sleuth/run1/01_kallisto_results/DB_103/run_info.json diff --git a/runs/run1/01_kallisto_results/DB_161/abundance.tsv b/runs/kallisto_sleuth/run1/01_kallisto_results/DB_161/abundance.tsv similarity index 100% rename from runs/run1/01_kallisto_results/DB_161/abundance.tsv rename to runs/kallisto_sleuth/run1/01_kallisto_results/DB_161/abundance.tsv diff --git a/runs/run1/01_kallisto_results/DB_161/run_info.json b/runs/kallisto_sleuth/run1/01_kallisto_results/DB_161/run_info.json similarity index 100% rename from runs/run1/01_kallisto_results/DB_161/run_info.json rename to runs/kallisto_sleuth/run1/01_kallisto_results/DB_161/run_info.json diff --git a/runs/run1/01_kallisto_results/DB_163/abundance.tsv b/runs/kallisto_sleuth/run1/01_kallisto_results/DB_163/abundance.tsv similarity index 100% rename from runs/run1/01_kallisto_results/DB_163/abundance.tsv rename to runs/kallisto_sleuth/run1/01_kallisto_results/DB_163/abundance.tsv diff --git a/runs/run1/01_kallisto_results/DB_163/run_info.json b/runs/kallisto_sleuth/run1/01_kallisto_results/DB_163/run_info.json similarity index 100% rename from runs/run1/01_kallisto_results/DB_163/run_info.json rename to runs/kallisto_sleuth/run1/01_kallisto_results/DB_163/run_info.json diff --git a/runs/run1/01_kallisto_results/DB_165/abundance.tsv b/runs/kallisto_sleuth/run1/01_kallisto_results/DB_165/abundance.tsv similarity index 100% rename from runs/run1/01_kallisto_results/DB_165/abundance.tsv rename to runs/kallisto_sleuth/run1/01_kallisto_results/DB_165/abundance.tsv diff --git a/runs/run1/01_kallisto_results/DB_165/run_info.json b/runs/kallisto_sleuth/run1/01_kallisto_results/DB_165/run_info.json similarity index 100% rename from runs/run1/01_kallisto_results/DB_165/run_info.json rename to runs/kallisto_sleuth/run1/01_kallisto_results/DB_165/run_info.json diff --git a/runs/run1/03_kallisto_mappingStats.csv b/runs/kallisto_sleuth/run1/03_kallisto_mappingStats.csv similarity index 100% rename from runs/run1/03_kallisto_mappingStats.csv rename to runs/kallisto_sleuth/run1/03_kallisto_mappingStats.csv diff --git a/runs/run1/03_kallisto_tpmMatrix.csv b/runs/kallisto_sleuth/run1/03_kallisto_tpmMatrix.csv similarity index 100% rename from runs/run1/03_kallisto_tpmMatrix.csv rename to runs/kallisto_sleuth/run1/03_kallisto_tpmMatrix.csv diff --git a/runs/run1/04_sleuth_dge.csv b/runs/kallisto_sleuth/run1/04_sleuth_dge.csv similarity index 100% rename from runs/run1/04_sleuth_dge.csv rename to runs/kallisto_sleuth/run1/04_sleuth_dge.csv diff --git a/runs/run1/05_shinyPrep.RData b/runs/kallisto_sleuth/run1/05_shinyPrep.RData similarity index 100% rename from runs/run1/05_shinyPrep.RData rename to runs/kallisto_sleuth/run1/05_shinyPrep.RData diff --git a/runs/run1/isa.run.xlsx b/runs/kallisto_sleuth/run1/isa.run.xlsx similarity index 100% rename from runs/run1/isa.run.xlsx rename to runs/kallisto_sleuth/run1/isa.run.xlsx diff --git a/runs/kallisto_sleuth/run2/03_kallisto_mappingStats.csv b/runs/kallisto_sleuth/run2/03_kallisto_mappingStats.csv new file mode 100644 index 0000000..21b8d2b --- /dev/null +++ b/runs/kallisto_sleuth/run2/03_kallisto_mappingStats.csv @@ -0,0 +1,7 @@ +"ID","n_targets","n_bootstraps","n_processed","n_pseudoaligned","n_unique","p_pseudoaligned","p_unique","kallisto_version","index_version","start_time","call" +"DB_097",31434,100,11071656,7663654,7522185,69.2,67.9,"0.44.0",10,"Fri Aug 6 15:48:13 2021","kallisto quant --single -b 100 -t 30 -l 200 -s 20 -i /Users/dominikbrilhaus/sciebo/CEPLAS_DM/CEPLAS_ARCs/ARC0005//runs/01_kallisto_index -o /Users/dominikbrilhaus/sciebo/CEPLAS_DM/CEPLAS_ARCs/ARC0005//runs/01_kallisto_results/DB_097 /Users/dominikbrilhaus/sciebo/CEPLAS_DM/CEPLAS_ARCs/ARC0005/assays/Talinum_RNASeq_minimal/dataset/DB_097_CAMMD_CAGATC_L001_R1_001.fastq.gz" +"DB_099",31434,100,14332278,10066506,9848811,70.2,68.7,"0.44.0",10,"Fri Aug 6 15:49:19 2021","kallisto quant --single -b 100 -t 30 -l 200 -s 20 -i /Users/dominikbrilhaus/sciebo/CEPLAS_DM/CEPLAS_ARCs/ARC0005//runs/01_kallisto_index -o /Users/dominikbrilhaus/sciebo/CEPLAS_DM/CEPLAS_ARCs/ARC0005//runs/01_kallisto_results/DB_099 /Users/dominikbrilhaus/sciebo/CEPLAS_DM/CEPLAS_ARCs/ARC0005/assays/Talinum_RNASeq_minimal/dataset/DB_099_CAMMD_CTTGTA_L001_R1_001.fastq.gz" +"DB_103",31434,100,13083957,9007720,8828236,68.8,67.5,"0.44.0",10,"Fri Aug 6 15:50:41 2021","kallisto quant --single -b 100 -t 30 -l 200 -s 20 -i /Users/dominikbrilhaus/sciebo/CEPLAS_DM/CEPLAS_ARCs/ARC0005//runs/01_kallisto_index -o /Users/dominikbrilhaus/sciebo/CEPLAS_DM/CEPLAS_ARCs/ARC0005//runs/01_kallisto_results/DB_103 /Users/dominikbrilhaus/sciebo/CEPLAS_DM/CEPLAS_ARCs/ARC0005/assays/Talinum_RNASeq_minimal/dataset/DB_103_CAMMD_AGTCAA_L001_R1_001.fastq.gz" +"DB_161",31434,100,12785081,9058396,8740183,70.9,68.4,"0.44.0",10,"Fri Aug 6 15:51:58 2021","kallisto quant --single -b 100 -t 30 -l 200 -s 20 -i /Users/dominikbrilhaus/sciebo/CEPLAS_DM/CEPLAS_ARCs/ARC0005//runs/01_kallisto_index -o /Users/dominikbrilhaus/sciebo/CEPLAS_DM/CEPLAS_ARCs/ARC0005//runs/01_kallisto_results/DB_161 /Users/dominikbrilhaus/sciebo/CEPLAS_DM/CEPLAS_ARCs/ARC0005/assays/Talinum_RNASeq_minimal/dataset/DB_161_reC3MD_GTCCGC_L001_R1_001.fastq.gz" +"DB_163",31434,100,14312514,10054493,9707941,70.2,67.8,"0.44.0",10,"Fri Aug 6 15:53:12 2021","kallisto quant --single -b 100 -t 30 -l 200 -s 20 -i /Users/dominikbrilhaus/sciebo/CEPLAS_DM/CEPLAS_ARCs/ARC0005//runs/01_kallisto_index -o /Users/dominikbrilhaus/sciebo/CEPLAS_DM/CEPLAS_ARCs/ARC0005//runs/01_kallisto_results/DB_163 /Users/dominikbrilhaus/sciebo/CEPLAS_DM/CEPLAS_ARCs/ARC0005/assays/Talinum_RNASeq_minimal/dataset/DB_163_reC3MD_GTGAAA_L001_R1_001.fastq.gz" +"DB_165",31434,100,13679479,9552504,9264346,69.8,67.7,"0.44.0",10,"Fri Aug 6 15:54:35 2021","kallisto quant --single -b 100 -t 30 -l 200 -s 20 -i /Users/dominikbrilhaus/sciebo/CEPLAS_DM/CEPLAS_ARCs/ARC0005//runs/01_kallisto_index -o /Users/dominikbrilhaus/sciebo/CEPLAS_DM/CEPLAS_ARCs/ARC0005//runs/01_kallisto_results/DB_165 /Users/dominikbrilhaus/sciebo/CEPLAS_DM/CEPLAS_ARCs/ARC0005/assays/Talinum_RNASeq_minimal/dataset/DB_165_re-C3MD_GTGAAA_L002_R1_001.fastq.gz" diff --git a/runs/kallisto_sleuth/run2/03_kallisto_tpmMatrix.csv b/runs/kallisto_sleuth/run2/03_kallisto_tpmMatrix.csv new file mode 100644 index 0000000..b36c478 --- /dev/null +++ b/runs/kallisto_sleuth/run2/03_kallisto_tpmMatrix.csv @@ -0,0 +1,31435 @@ +"target_id","CAM_01","CAM_02","CAM_03","reC3_01","reC3_02","reC3_03" +"Tfru00001_t",2.38259718580395,2.52445339258139,2.99559686081831,4.485419618992,5.15554766862727,4.81121852299496 +"Tfru00002_t",25.1245986606198,25.2087869222372,26.5380313245026,30.658236957757,29.3913436146477,20.8234431654784 +"Tfru00003_t",0,0,0,0.309365430148544,0.725542630523583,1.05024822889448 +"Tfru00004_t",0.103844717888069,0.0833541471161738,0.0894261013194486,0.107415134520747,0.449405712721379,0.0759765881661035 +"Tfru00005_t",0.103461055137251,0.996554261334748,0.623669965236499,113.224757627604,148.648845382204,129.135350356906 +"Tfru00006_t",5.25064854821547,8.28385729734509,7.17220460021974,9.17681755934687,14.8875327806903,12.3195055255455 +"Tfru00007_t",3.06013893760537,4.10665056164903,3.74698927321371,6.23178959717579,6.24916931819361,7.38140445268395 +"Tfru00008_t",14.2043439781122,26.3981885595812,13.4442914669108,26.9366560835528,22.7611500493748,19.0527641655261 +"Tfru00009_t",6.98648923869327,9.55423377037359,10.3244912930937,13.6504343999739,14.7070095797832,16.21819340388 +"Tfru00010_t",244.725880160303,254.236879163414,239.055409283695,234.24905727355,255.175913290995,153.47176428698 +"Tfru00011_t",249.648439921802,235.855086169051,261.859652611709,239.044084421155,277.23708760388,237.068812271962 +"Tfru00012_t",0,0,0,0,0,0 +"Tfru00013_t",0,0,0,0,0,0 +"Tfru00014_t",0.0481534782380506,0.0385649628570616,0.040081234898993,0,0.0409396548817672,0.0683249248759905 +"Tfru00015_t",9.12352121874593,7.38645814357469,6.05895371227259,8.36401371837615,8.18030748581089,6.6074939313356 +"Tfru00016_t",0,0,0,0,0,0 +"Tfru00017_t",73.8872427283506,59.7748520926179,84.9711456238005,79.6773332599202,64.3547735725843,54.9950535288643 +"Tfru00018_t",1.17497030449515,1.88625188858599,2.02365639069639,1.82305266770431,1.01697684361232,3.00877915233859 +"Tfru00019_t",100.042891419037,88.5386478055069,90.5702092996186,107.46269236286,116.564177910843,73.6642320031186 +"Tfru00020_t",4.02402645039706,3.40000865631855,5.41073001773245,4.81959974168133,4.94942973931415,4.2870587946129 +"Tfru00021_t",83.6566488692519,95.2586986660982,83.0071898994768,27.350267688798,24.8757888457407,26.3329256308797 +"Tfru00022_t",0,0,0,0,0,0 +"Tfru00023_t",0,0,0,0.0722351154009016,0,0 +"Tfru00024_t",0,0,0,0,0,0 +"Tfru00025_t",10.3672379845616,10.3302390664317,10.2823272906906,11.7590775902453,8.66377719537418,6.800389597812 +"Tfru00026_t",1.8199540035594,2.42604099287411,1.93108487495278,3.59697341954331,3.99433883682836,2.59175687469059 +"Tfru00027_t",55.063462876232,42.4428339387146,49.6338143237525,83.0396302423057,82.6243070202033,67.3009460579787 +"Tfru00028_t",21.6026683126579,10.7893608027175,14.2624411478655,16.3866391217165,13.9195073409354,13.6979276184499 +"Tfru00029_t",3.29320676944074,2.53549978863862,2.9517052114713,38.5135675842526,37.1713172155247,30.1424653388556 +"Tfru00030_t",21.6360831128322,23.8598828656384,22.7863826973994,22.4807913845528,21.2574115640561,19.0384814079383 +"Tfru00031_t",86.4958908771116,87.5837535575628,99.1007884405973,14.1403444097653,15.9195990519397,11.2746559444835 +"Tfru00032_t",21.9121553586945,23.561903819911,19.2257316055188,25.4025163550232,23.2598419325332,12.7043922869481 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install.packages("BiocManager") +BiocManager::install() +BiocManager::install("devtools") # only if devtools not yet installed +BiocManager::install("pachterlab/sleuth") +library(sleuth) +BiocManager::install("pachterlab/sleuth") +library(sleuth) +BiocManager::install("pachterlab/sleuth") +install.packages('Matrix', dependencies = TRUE) +BiocManager::install("pachterlab/sleuth") +source("http://bioconductor.org/biocLite.R") +devtools::install_github("pachterlab/sleuth") +remotes::install_github("pachterlab/sleuth#260") +library(sleuth) +# BiocManager::install("pachterlab/sleuth") +# There's currently (early 2022) an issue with sleuth installation. +# Using this workaround from https://github.com/pachterlab/sleuth/issues/259#issuecomment-1001076030 +remotes::install_github("pachterlab/sleuth#260") +library(sleuth) +library(tidyverse) +library(jsonlite) +library(openxlsx) +?commandArgs +BiocManager::install() +library(sleuth) +required_packages <- +c('tidyverse', ## data wrangling and plotting +'jsonlite', ## read/write json files +'openxlsx') ## read/write xlsx files +for(package in required_packages) +{ +print(package) +## Check if package is installed. If not, install +if(!package %in% row.names(installed.packages())){install.packages(package, +repos ="https://cran.uni-muenster.de/")} +# ## Check if package is up to date. If not, update +# update.packages(package, repos = "https://cran.uni-muenster.de/") +## Load package +library(package, character.only = T) +} +for(package in required_packages) +{ +## Check if package is installed. If not, install +if(!package %in% row.names(installed.packages())){install.packages(package, +repos ="https://cran.uni-muenster.de/")} +# ## Check if package is up to date. If not, update +# update.packages(package, repos = "https://cran.uni-muenster.de/") +## Load package +library(package, character.only = T) +print(paste(package, "installed")) +} +{ +## Check if package is installed. If not, install +if(!package %in% row.names(installed.packages())){install.packages(package, +repos ="https://cran.uni-muenster.de/")} +# ## Check if package is up to date. If not, update +# update.packages(package, repos = "https://cran.uni-muenster.de/") +## Load package +library(package, character.only = T) +print(paste("installed R package", package)) +} +for(package in required_packages) +{ +## Check if package is installed. If not, install +if(!package %in% row.names(installed.packages())){install.packages(package, +repos ="https://cran.uni-muenster.de/")} +# ## Check if package is up to date. If not, update +# update.packages(package, repos = "https://cran.uni-muenster.de/") +## Load package +library(package, character.only = T) +print(paste("installed R package", package)) +} +run_name="run1" +arc_root="~/03DataPLANT_gitlab/samplearc_rnaseq/" +# Load the sleuth object +load(file = paste0(arc_root, 'runs/', run_name, '/03_kallisto_sleuthObject.RData')) +so <- sleuth_fit(so) +so <- sleuth_fit(so, ~Group, 'full') +so <- sleuth_fit(so, ~Photosynthesis.mode, 'full') +so <- sleuth_fit(so, ~1, 'reduced') +so <- sleuth_lrt(so, 'reduced', 'full') +sleuth_table <- sleuth_results(so, 'reduced:full', 'lrt', show_all = FALSE) +write.csv(sleuth_table, paste0(arc_root, 'runs/', run_name, '/04_sleuth_dge.csv'), row.names = F) +library(openxlsx) +expression_data <- read.csv(file = paste0(arc_root, 'runs/', run_name, '/03_kallisto_df.csv')) +available_genes <- unique(expression_data$target_id) +save(expression_data, available_genes, file = paste0(arc_root, 'runs/', run_name, '/05_shinyPrep.RData')) +load(file = "../runs/run1/05_shinyPrep.RData") +required.packages <- c('knitr', 'kableExtra', ## RMarkdown, +"shiny", "tidyverse", ## data loading and shaping +"RColorBrewer", "shiny" ## plotting +) +for(package in required.packages) +{ +print(package) +## Check if package is installed. If not, install +if(!package %in% row.names(installed.packages())) +{install.packages(package, repos ="https://cran.uni-muenster.de/")} +## Load package +library(package, character.only = T) +} +required.packages <- c('kableExtra', ## RMarkdown, +"shiny", "tidyverse", ## data loading and shaping +"RColorBrewer", "shiny" ## plotting +) +for(package in required.packages) +{ +print(package) +## Check if package is installed. If not, install +if(!package %in% row.names(installed.packages())) +{install.packages(package, repos ="https://cran.uni-muenster.de/")} +## Load package +library(package, character.only = T) +} +current_selection <- sample(expression_data$target_id, 10) +plot_set <- subset(expression_data, target_id %in% current_selection) +ggplot(plot_set, aes(x = Group, y = tpm, group = Group)) + +stat_summary(fun = 'mean', geom = 'bar') + +geom_point(size = 0.5) + +facet_wrap(~ target_id , scales = "free") + +theme_minimal() +plot_set <- subset(expression_data, target_id %in% current_selection) +ggplot(plot_set, aes(x = Photosynthesis.mode, y = tpm, group = Photosynthesis.mode)) + +stat_summary(fun = 'mean', geom = 'bar') + +geom_point(size = 0.5) + +facet_wrap(~ target_id , scales = "free") + +theme_minimal() diff --git a/workflows/kallisto_sleuth/01_KallistoIndex.sh b/workflows/kallisto_sleuth/01_KallistoIndex.sh new file mode 100755 index 0000000..ec37c24 --- /dev/null +++ b/workflows/kallisto_sleuth/01_KallistoIndex.sh @@ -0,0 +1,25 @@ + +run_name=$1 +arc_root=$2 + +## Manual: http://pachterlab.github.io/kallisto/manual.html + +## MacOS Installation +# ruby -e "$(curl -fsSL https://raw.githubusercontent.com/Homebrew/install/master/install)" +# brew install kallisto + + +# git clone https://github.com/pachterlab/kallisto.git +# cd kallisto +# mkdir build +# cd build +# cmake .. -DUSE_HDF5=ON + + +kallisto version +kallisto cite + +### Build index + +kall_ref=$arc_root'/studies/TalinumGenomeDraft/resources/Talinum.gm.CDS.nt.fa' +kallisto index -i $arc_root'/runs/'$run_name'/01_kallisto_index' $kall_ref \ No newline at end of file diff --git a/workflows/kallisto_sleuth/01_KallistoQuant.sh b/workflows/kallisto_sleuth/01_KallistoQuant.sh old mode 100644 new mode 100755 index bca3fd8..f65e0ac --- a/workflows/kallisto_sleuth/01_KallistoQuant.sh +++ b/workflows/kallisto_sleuth/01_KallistoQuant.sh @@ -1,32 +1,26 @@ -#!/usr/bin/bash +#!/usr/bin/env bash + # Map RNASeq reads via kallisto run_name=$1 arc_root=$2 -## Manual: http://pachterlab.github.io/kallisto/manual.html - -kallisto version -kallisto cite - -### Build index - -kall_ref=$arc_root'/studies/TalinumGenomeDraft/resources/Talinum.gm.CDS.nt.fa' -kallisto index -i $arc_root'/runs/'$run_name'/01_kallisto_index' $kall_ref - ### Align reads -ILLUMINASAMPLES=$(ls ${arc_root}'/assays/Talinum_RNASeq_minimal/dataset/'*fastq.gz) +ILLUMINASAMPLES=$(ls ${arc_root}"/assays/Talinum_RNASeq_minimal/dataset/"*"fastq.gz") -mkdir $arc_root'/runs/'$run_name'/01_kallisto_results/' +mkdir -p $arc_root'/runs/'$run_name'/01_kallisto_results/' for j in $ILLUMINASAMPLES; do sampleName=$(echo $j | sed -e 's|.*/||' | cut -c -6) # cut away path. retain only first six chars of file name echo $sampleName - kallisto quant --single -b 100 -t 30 -l 200 -s 20 -i $arc_root'/runs/'$run_name'/01_kallisto_index' -o $arc_root'/runs/'$run_name'/01_kallisto_results/'$sampleName $j + kallisto quant --single -b 100 -t 8 -l 200 -s 20 -i $arc_root'/runs/'$run_name'/01_kallisto_index' -o $arc_root'/runs/'$run_name'/01_kallisto_results/'$sampleName $j echo 'Kallisto done' -done \ No newline at end of file +done + + + diff --git a/workflows/kallisto_sleuth/02_InstallPackages.R b/workflows/kallisto_sleuth/02_InstallPackages.R index f328f61..163e75e 100644 --- a/workflows/kallisto_sleuth/02_InstallPackages.R +++ b/workflows/kallisto_sleuth/02_InstallPackages.R @@ -12,13 +12,15 @@ ### sleuth installation -if (!requireNamespace("BiocManager", quietly = TRUE)) install.packages("BiocManager") -BiocManager::install() -BiocManager::install("devtools") # only if devtools not yet installed +if (!requireNamespace("BiocManager", quietly = TRUE)){install.packages("BiocManager")} +if(!"BiocManager" %in% row.names(installed.packages())){BiocManager::install()} +if(!"devtools" %in% row.names(installed.packages())){BiocManager::install("devtools")} + # BiocManager::install("pachterlab/sleuth") # There's currently (early 2022) an issue with sleuth installation. # Using this workaround from https://github.com/pachterlab/sleuth/issues/259#issuecomment-1001076030 -remotes::install_github("pachterlab/sleuth#260") + +if(!"sleuth" %in% row.names(installed.packages())){remotes::install_github("pachterlab/sleuth#260")} library(sleuth) diff --git a/workflows/kallisto_sleuth/03_KallistoCollect.R b/workflows/kallisto_sleuth/03_KallistoCollect.R index ff06926..f36e8f7 100644 --- a/workflows/kallisto_sleuth/03_KallistoCollect.R +++ b/workflows/kallisto_sleuth/03_KallistoCollect.R @@ -45,7 +45,12 @@ assay_data <- merge(study_data, assay_data, by.x = "Sample.Name", by.y = "Sourc assay_data <- assay_data[, !apply(assay_data, 2, function(x){sum(is.na(x)) == nrow(assay_data)})] # Pointer to kallisto results folder -base_dir <- paste0(arc_root, 'runs/', run_name, '/01_kallisto_results/') +# base_dir <- paste0(arc_root, 'runs/', run_name, '/01_kallisto_results/') + +######################## +### Note: hard coded link due to issues with kallisto version +base_dir <- paste0(arc_root, 'runs/kallisto_sleuth/run1/01_kallisto_results') +######################## # A list of paths to the kallisto results indexed by the sample IDs is collated with kal_dirs <- dir(base_dir, full.names = T) ## Sleuth requires full paths @@ -58,7 +63,7 @@ s2c$path <- kal_dirs s2c <- s2c[order(s2c$sample), ] # Build a sleuth object -so <- sleuth_prep(s2c, ~Photosynthesis.mode) +so <- sleuth_prep(s2c, ~Photosynthesis.mode, num_cores = 1) save(so, file = paste0(arc_root, 'runs/', run_name, '/03_kallisto_sleuthObject.RData')) # Extract expression tables diff --git a/workflows/kallisto_sleuth_wrapper.sh b/workflows/kallisto_sleuth_wrapper.sh index d0bbe31..641796b 100755 --- a/workflows/kallisto_sleuth_wrapper.sh +++ b/workflows/kallisto_sleuth_wrapper.sh @@ -1,4 +1,4 @@ - #!/usr/bin/bash +#!/usr/bin/env bash ####################################### #### To be replaced by CWL routine #### @@ -8,21 +8,30 @@ # Execute within <ARC root>/workflows: ## change rights: `chmod a+x kallisto_sleuth_wrapper.sh` ## execute script: `./kallisto_sleuth_wrapper.sh <run_name> <arc_root>` -## Example: ./kallisto_sleuth_wrapper.sh "run2" "~/03DataPLANT_gitlab/samplearc_rnaseq/" +## Example: ./kallisto_sleuth_wrapper.sh "run2" "/Users/dominikbrilhaus/03DataPLANT_gitlab/samplearc_rnaseq/" run_name='kallisto_sleuth/'$1 arc_root=$2 -mkdir $arc_root'/workflows/'$run_name +# Note: due to some not well documented change in kallisto development (use of hd5) the kallisto quant part is currently not reproducible and commented out +# Pointer to `runs/kallisto_sleuth/run1/01_kallisto_results` is thus hard coded into 03_KallistoCollect.R -# chmod a+x 01_KallistoQuant.sh -# ./01_KallistoQuant.sh $run_name $arc_root +mkdir -p $arc_root'/runs/'$run_name -# Rscript 02_InstallPackages.R -# Rscript 03_KallistoCollect.R $run_name $arc_root -# Rscript 04_Sleuth.R $run_name $arc_root -# Rscript 05_plot_shinyPrep.R $run_name $arc_root +# chmod a+x ./kallisto_sleuth/01_KallistoIndex.sh +# ./kallisto_sleuth/01_KallistoIndex.sh $run_name $arc_root +# chmod a+x kallisto_sleuth/01_KallistoQuant.sh +# ./kallisto_sleuth/01_KallistoQuant.sh $run_name $arc_root + +Rscript ./kallisto_sleuth/02_InstallPackages.R +echo "r insallations done" +Rscript ./kallisto_sleuth/03_KallistoCollect.R $run_name $arc_root +echo "kallisto collect done" +Rscript ./kallisto_sleuth/04_Sleuth.R $run_name $arc_root +echo "sleuth done" +Rscript ./kallisto_sleuth/05_plot_shinyPrep.R $run_name $arc_root +echo "shiny prep done" # store variables for test runs in R -# run_name="run1" -# arc_root="~/03DataPLANT_gitlab/samplearc_rnaseq/" \ No newline at end of file +# run_name="kallisto_sleuth/run2" +# arc_root="/Users/dominikbrilhaus/03DataPLANT_gitlab/samplearc_rnaseq/" \ No newline at end of file diff --git a/workflows/plot_shinyApp.Rmd b/workflows/plot_shinyApp.Rmd index 19b2916..a710642 100644 --- a/workflows/plot_shinyApp.Rmd +++ b/workflows/plot_shinyApp.Rmd @@ -14,7 +14,7 @@ knitr::opts_chunk$set(echo = TRUE) # Load data -load(file = "../runs/run1/05_shinyPrep.RData") +load(file = "../runs/kallisto_sleuth/run2/05_shinyPrep.RData") ``` -- GitLab