alias: GSM2735454
title: GSM2735454: RNA-seq GFP rep 3; Mus musculus; ssRNA-seq
library_construction_protocol: Cells were lysed directly in plates and total RNA was extracted using TRIzol reagent (Invitrogen) according to manufacturer's instructions. Strand-specific libraries were generated using the TruSeq Stranded mRNA Library Prep Kit (Illumina). Polyadenylated RNA from 1000 ng intact total RNA was purified using oligo-dT beads. Following purification, the RNA was fragmented, random primed and reverse transcribed using SuperScript II Reverse Transcriptase (Invitrogen). Second strand synthesis was performed using Polymerase I and RNase H with replacement of dTTP for dUTP. The generated cDNA fragments were 3' end adenylated and ligated to Illumina Paired-end sequencing adapters and subsequently amplified by 12 cycles of PCR. The libraries were analyzed on a 2100 Bioanalyzer using a 7500 chip (Agilent), diluted, and pooled in equimolar ratio into a 13-plex, 10 nM sequencing pool.
identifiers.namespace: GEO
identifiers.id: GSM2735454