GSM3258245: HFD Control; Mus musculus; RNA-Seq
1 μg total RNA with an RNA integrity number (RIN) of 8.0 was used for mRNA purification using oligo-dT beads (TruSeq RNA Sample Preparation Kit, Illumina). The purified mRNA was fragmented and reverse transcribed using Superscript II Reverse Transcriptase (Life Technologies). Second strand cDNA was synthesized on the first strand template using RNaseH and DNA polymerase I. The cDNAs were cleaned using Beckman Coulter Agencourt Ampure XP SPRI beads. The sequencing library was prepared by random fragmentation of the cDNA sample, followed by 5' and 3' adapter ligation, after end-repair and the addition of an 'A' base and SPRI cleanup. Adapter-ligated fragments were enriched by PCR. The individual libraries were quantified using a NanoDrop spectrophotometer (Thermo Scientific) and validated for quality with a Bioanalyzer (Agilent Technologies). Adapter-ligated fragments are then PCR amplified and gel purified
GEO Accession
GSM3258245