title: GSM1399379: T 40 NU2; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was extracted from D1 mouse cell line with Trizol Reagent (Invitrogen) followed by Rneasy micro clean-up procedure (Qiagen) as per manufacturer's instructions. Total RNA integrity was assessed by Agilent 2100 Bioanalyzer (Agilent) TruSeq: D1 total RNA was mixed with ERCC RNA Spike in Control Mix 1 (Ambion) and subjected to library preparation using Illumina TruSeq RNA Sample Preparation kit version 1 (Low Sample Protocol) with slight modifications. Briefly, polyA-containing RNA molecules were purified from the D1 RNA samples using polyT oligo-attached magnetic beads. Thermal fragmentation followed after two rounds of enrichment for polyA+ mRNA. cDNA was synthesized from the RNA fragments using reverse transcriptase (Superscript II) and random primers. This was followed by second strand cDNA synthesis and the resulting dsDNA went through an end repair process, the addition of a single A base, and ligation of the adapters. The products were purified and enriched with 12 cycles of PCR to create the cDNA library. Two additional rounds of purification of the cDNA libraries were done using Agencourt Ampure XP SPRI beads (Beckamn Courter) to remove >600 bp double stranded cDNA. The length distribution of cDNA libraries was monitored using DNA 1000 kits on the Agilent Bioanalyzer
identifiers.namespace: GEO
identifiers.id: GSM1399379