RNA-seq of Homo sapiens: adult male prostate adenocarcinoma
Total RNA was isolated from 33 tumor tissue samples using the MagNA Pure Compact RNA Isolation Kit (Roche, Switzerland). RNA concentration was determined using a NanoDrop 100 spectrophotometer (PeqLab, Germany) and Qubit 2.0 Fluorometer (Life Technologies, USA). The RIN (RNA Integrity Number) parameter, which characterizes the integrity of RNA, was evaluated using the Agilent 2100 Bioanalyzer (Agilent Technologies, USA). RIN for all samples studied was no less than 8.0. Libraries were prepared using the TruSeq Stranded mRNA LT kit (Illumina, USA) according to the manufacturer's protocol. The average size of prepared cDNA libraries was 300 bp. Library selection was PolyA selection, enrichment for messenger RNA (mRNA). Libraries were further validated prior to sequencing using quantitative PCR (PCR-RT). Sequencing was performed on the NextSeq500 System (Illumina, USA).