title: GSM3904188: S1: Wild-type, pair-fed replicate 1; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was isolated using Trizol, DNase digested, and then cleaned-up using the GeneJet system from ThermoFisher. Poly A purification and RNA fragmentation were performed on 600ng total RNA. Superscript II was used to generate cDNA and then RNA template was removed. Second strand synthesis was performed with incorporation of dUTP to ensure stranded libraries, and ds cDNA was purified with AMPure XP beads. 3' ends were adenylated, indexing adapters were ligated onto the ends, and libraries once again purified with AMPure beads. 15 cycles of PCR were used to enrich DNA fragments, followed by two AMPure bead clean-up steps. Libraries were loaded onto an Agilent DNA 1000 chip and validated on an Agilent 2100 Bioanalyzer. Quantitation was performed with the Illumina Library Quantification Kit, ABI Prism qPCR Mix from Kapa Biosystems.