GSM2610696: RNA activated CD8+ T cells (Nfatc2ko); Mus musculus; RNA-Seq
total RNA from each sample was isolated by using Trizol and quality of RNA was assessed on a RNA6000 Nano Chip and a Bioanalyzer2100 (Agilent) Barcoded mRNA-seq cDNA libraries were prepared from 500ng of total RNA (RIN>8) using NEBNext® Poly(A) mRNA Magnetic Isolation Module and NEBNext® Ultra™ RNA Library Prep Kit for Illumina® according to the manual. All quality controls were done using Invitrogen’s Qubit HS assay and fragment size was determined using Agilent’s 2100 Bioanalyzer HS DNA assay. Barcoded RNA-Seq libraries were onboard clustered usingHiSeq® Rapid SR Cluster Kit v2 using 8pM and 59bps were sequenced on the Illumina HiSeq2500 using HiSeq® Rapid SBS Kit v2 (50 Cycle). The raw output data of the HiSeq was preprocessed according to the Illumina standard protocol.