Transcriptomics analysis of gene expression in multiple human and mouse cells and tissues

RNA was isolated from siCTRL, siNSUN2 HeLa or 3T3-L1 cells, human cells (embryonic kidney T cell line 293T; colon cancer cell line HCT116; hepatocellular carcinoma cell line Huh7; embryonic lung fibroblast cell line MRC-5; glioma cell line SNB19), mouse cells (kidney collecting duct cell line M-1; cervical cancer cell line U14) and multiple mouse tissues using the TRIzol (Invitrogen) reagent by following the company manual. For all samples the RNA integrity was checked using an Agilent Bioanalyzer 2100. All samples showed a RIN (RNA integrity number) of higher than 9. Approximately 2.5 µg of total RNA was then used for library preparation using a TruSeq™ RNA Sample Prep Kit v2 (Illumina, San Diego, CA, USA) according to the manufacturer's protocol.The libraries were sequenced using HiSeq2000 (Illumina) in paired-read mode, creating reads with a length of 101 bp. Sequencing chemistry v2 (Illumina) was used and samples were multiplexed in two samples per lane. Overall design: Examination of gene expressive levels in mammalian transcriptome