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rnaseq-ps23

RNA-seq pipeline, from raw reads to biological interpretaion of time-course experiment. Whenewer possible bash command-line tools are preffered over R.

  • English localization

alternatywy.r - just some links to web pages with potentially usefu information.

Archival deprecated analyses

Directory deprecated

prb.masigpro.r - test of time-course analysis in maSigPro R package.

see.gen2plot.r - modification of the see.genes function, used to prevent over-writing of plots in R (used from command-line) In the end this package will not be used as it don't allow to compare multiple groups and the contact with developers is poor.

Analyses on simulated or test data

Directory test-simulated-data

The first trials of read-processing are given in rnaseq_pipeline.sh

deseq-analysis.r - test of differential gene expression analysis in DESeq2 package in R. Two versions, two replications and three replications

go-enrich.r - GO over and underrepresentation test in goseq

mfuzz-analysis.r - fuzzy (soft) clustering, use data from deseq-analysis.r. Here also corrected script colorbar.R for legend is used.

  • other explorative analyses

Analyses on real data

Directory sequence-analysis rnaseq-ost.sh - the clean read-processing workflow, based on rnaseq_pipeline.sh

rnaseq72.sh - the clean read-processing workflow for whole first replication of experiment

counts4r.sh - construction of files used for DESeqDataSet in DESeq2 package in R

bam4labels.sh - commands used to change header of the counts file to match sample labels

Directory stats-and-visuals

boxplot-r.r - simple boxplot of raw counts

counts-boxplots.r - more advanced boxplots of read counts, involving transformation and FPKM calculation

explorative-plots.r - exploring patterns in count data from first sequencing results (PCA and clustering)

overrepresented-seqs-check.sh - what are overrepresented sequences in FastQC results?

multiplot.r - script for plotting multiple PCA plots (plotPCA command) at the same area

  • correct data after new sequencing of "U3" sample - we have results
  • full analysis of 2nd replication, plus joint PCA/clustering analysis
  • update sample names in count file to include replication number
  • check, if it is possible to make DE analysis on two replications (DEseq2) - OK

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