# TODO define project name export project="TODO define project name" # screen -R ${project} ## madmax if [ "$HOSTNAME"=="falcon1" ]; then export baseDir="/projects/bioinfo/holger/projects/${project}" export PROJECT_SCRIPTS="/projects/bioinfo/holger/scripts/${project}" export NGS_TOOLS="/projects/bioinfo/scripts/ngs_tools/dev" fi ## bioinfo if [ $(hostname) == "bioinformatics-srv1" ]; then export baseDir=/net/mack/lustre/projects/bioinfo/holger/projects/${project} export PROJECT_SCRIPTS==/net/mack/lustre/projects/bioinfo/holger/scripts/${project} export NGS_TOOLS=/net/mack/lustre/projects/bioinfo/scripts/ngs_tools/dev fi source ${NGS_TOOLS}/dge_workflow/dge_utils.sh export PATH=${NGS_TOOLS}/dge_workflow:$PATH ## TODO define igenome to be used ## igenome=/projects/bioinfo/igenomes/Canis_familiaris/Ensembl/CanFam3.1 #igenome=<<<>>> ######################################################################################################################## ### Fetch the data mcdir ${baseDir}/originals wget -nc --user="USER" --password="PW" -r --no-directories --no-check-certificate -A "*fastq.gz" https:/projects.biotec.tu-dresden.de/ngs-filesharing/martaf/ mailme "$project: fastq download done" ### Basic QC dge_fastqc $(ls *fastq.gz) & ## todo make sure to also copy the sample sheet in here ######################################################################################################################## ### Apply renaming and merge lane replicates (but keep technical ones) ## todo adjust renaming scheme to project specifics mcdir $baseDir/lanereps_pooled echo ' devtools::source_url("https://raw.githubusercontent.com/holgerbrandl/datautils/v1.36/R/core_commons.R") sheetFile <- "../originals/natalied-FC_SN678_338-2015-5-12.xls" sampleSheet <- read_excel(sheetFile, "Fastqfiles") %>% select(File, SampleName) %>% mutate( bio_replicate=str_match(SampleName, "(.).*")[,2], sample = str_replace(SampleName, "[0-9]*", "") %>% str_replace_all(c("NC" = "no_culture", "NA" = "no_hormone", "ECD"="ecdysone_", "INS"="insulin_", "9"="9h", "4"="4h")), bio_sample=paste(sample, bio_replicate, sep="_") ) write_tsv(sampleSheet, path="renaming_scheme.txt") require(ggplot2) ggplot(sampleSheet, aes(bio_sample)) + geom_bar() + coord_flip() sampleSheet %>% group_by(bio_sample) %>% summarise( zcat=paste("zcat", paste(paste0("../originals/", File), collapse=" "), "| gzip -c >", paste0(bio_sample[1], ".fastq.gz")) ) %$% zcat %>% write_lines("lane_merge.cmd") ' | R --vanilla -q cat lane_merge.cmd | while read line; do # eval ${line} jl submit -j .repmerge "$line" done jl wait --email --report dge_fastqc $(ls *fastq.gz) & ######################################################################################################################## ### Alignment the reads mcdir $baseDir/alignments star_align.kts ${igenome} $(ls ${baseDir}/lanereps_pooled/*.fastq.gz) #dge_bam_correlate . & # part of star_align.kts now mailme "$project: mapping done" ######################################################################################################################## ### Differential Expression Analysis mcdir $baseDir/dge_analysis rend.R -e ${NGS_TOOLS}/dge_workflow/featcounts_deseq.R ../alignments/star_counts_matrix.txt 2>&1 | tee featcounts_deseq.log ## or use just some contrasts of interest #echo " #sample_1, sample_2 #unpolarised,liver_polar_stage3 #" | csvformat -T > contrasts.txt # #rend.R -e $NGS_TOOLS/dge_workflow/featcounts_deseq.R --contrasts contrasts.txt ../alignments/star_counts_matrix.txt 2>&1 | tee featcounts_deseq.log ## Term enrichment mcdir ${baseDir}/dge_analysis/dge_enrichment_analysis $NGS_TOOLS/common/cp_enrichment.R --overlay_expr_data ../plot_score_matrix.txt ../degs_by_contrast.txt contrast rend.R -e ${NGS_TOOLS}/common/cp_enrichment.R --overlay_expr_data ../plot_score_matrix.txt ../degs_by_contrast.txt contrast ######################################################################################################################## ### Sync back to project space ## bidirectional sync with project space ## todo define mount path on bioinfo for bidirectional synching ## ~/bin/unison $baseDir ssh://bioinfo///home/brandl/mnt/<> -fastcheck true -times -perms 0 # or use a uni-directional sync #rsync -avsn --delete ${baseDir} brandl@fileserver:/projects//file/server/path mailme "$project: sync done"