title: GSM1133714: Treatment Rat 5; Rattus norvegicus; RNA-Seq
library_construction_protocol: Livers were snap frozen using a dry ice and isopentane bath. Total RNA was extracted using RNA Stat-60 (Amsbio) following manufacturer's directions. We used the Illumina TruSeq RNA kit for library preparation with some modifications to the manufacturer’s directions (detailed below). Poly-A RNA was isolated by annealing biotinylated oligo-dT to total RNA and purifying using streptavidin-conjugated beads. RNA was fragmented for 4 minutes at 94°C to increase the population of 200-600 nt fragments. cDNA synthesis was accomplished using random hexamers. Fragment ends were repaired and 3’ ends were adenylated for adaptor ligation. Adaptor ligated fragments were enriched following 10 cycles of PCR. 320-600 bp molecules were size selected from the resulting amplicon pool using a PippinPrep (Sage Science). The final cDNA library was titrated by qRT-PCR and sequenced in 101 nt paired-end reads.
attributes.tag: GEO Accession
attributes.value: GSM1133714