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  • Use bedtools to count reads in features.
  • Converting mapped reads to feature counts.

The data files for this example are in the path:

Code Block

/corral-repl/utexas/BioITeam/ngs_course/ecoli_rnaseq

File Name

Description

SRR030252.fastq.gz

Illumina reads, 0K generation individual clone from population

SRR032374.fastq.gz

Illumina reads, 20K generation mixed population

SRR032376.fastq.gz

Illumina reads, 40K generation mixed population

NC_012967.1.fasta.gz

E. coli B str. REL606 genome

hg3. Using DESeq

Code Block
titleDESeq installation

login1$ R
...
>source("http://bioconductor.org/biocLite.R")
>biocLite("DESeq")
Code Block
titleUsing DESeq

login1$ R
...
> library("DESeq")
> combined = read.csv("combined.csv", header=T, row.names=1)
> design <- data.frame(
  row.names = colnames( combined ),
  condition = c( "Anc", "Anc", "EL", "EL", "EW", "EW"),
  libType = c( "single-end", "single-end", "single-end",
  "single-end", "single-end", "single-end" ) )
> design
> conds <- factor(design$condition)
> cds <- newCountDataSet( combined, conds )
> cds <- estimateSizeFactors( cds )
> sizeFactors( cds )
> res <- nbinomTest( cds, "EL", "EW" )
> write.csv(res, "EL-vs-EW.csv")