GSM2211867: 4h_Trans_A; Mus musculus; RNA-Seq
Total RNA was extracted using a Qiagen RNeasy mini kit. Libraries were prepared according to TruSeq RNA Sample Prep Kit v2. The first step was to purify the poly-A–containing mRNA molecules using poly-T oligo-attached magnetic beads. The mRNA was then fragmented into 120–210 bp pieces by incubating at 94 °C for 8 minutes. The cleaved RNA fragments were copied into first strand cDNA using reverse transcriptase and random hexamers. Second strand cDNA synthesis utilized DNA Polymerase I and RNase H. These fragments then went through an end repair process to blunt the ends. A single “A” base was added to the 3' end of the fragment, and then indexed adapters were ligated to the ends. Each adapter had a 7 bp sequence that was unique to one of the fifteen samples. The products were purified and amplified with PCR. This cDNA was quantified using an Agilent Technologies 2100 Bioanalyzer and then subjected to high-throughput sequencing by Illumina HiSeq 2000.
GEO Accession
GSM2211867