Illumina RNA-Seq of human male prostate cancer tumor
The samples were prepared in accordance with the Illumina RNA sample preparation protocol (Part # 1004898 Rev. A September 2008). Briefly, mRNAs were fragmented at elevated temperature using divalent cations and transcribed into cDNA, thereby generating a library of cDNA fragments. RNA-Seq adapters were then ligated to the blunt ends of the cDNA fragments. The library of cDNA fragments subsequently underwent a size-selection step in which cDNAs were first electrophoresed through a 2.5% agarose gel in TAE buffer. Then, the desired fragment size products (200 or 300 nt) were retrieved from the gel and subjected to PCR amplification using universal primer sites present at the end of the ligated adapters. The library was then subjected to quality control steps such as verification of fragment size and concentration measurements using the DNA 1000 Kit (Agilent Technologies) on an Agilent 2100 Bioanalyzer.