title: Illumina HiSeq 2500 paired end sequencing; RNA-seq of livers from glucagon receptor knock-out (GcgrKO) mice compared to wildtype littermates
design: RNA-seq of livers from glucagon receptor knock-out (GcgrKO) mice compared to wildtype littermates
library_name: GR413_p
library_construction_protocol: Mice were subject to a short-term fast of 4 hours before anesthesia with 2.5% isoflurane. A liver biopsy was taken from the right medial lobe and stored in RNAlater a -20 degree C until analysis. Raw sequencing data were processed using CASAVA v. 1.8 conversion software (Illumina). Total RNA was purified by an AllPrep DNA/RNA/ protein extraction kit (QIAgen) according to manufacturer's instructions and qualified using a 2100 Bioanalyzer instrument (Agilent Genomics). RNA-sequencing was done using TruSeq Stranded Total RNA Library Prep Kit (Illumina) according to the manufacturer's protocol: Ribosomal RNA was depleted using TruSeq Standed RNA Ribominus Gold (Illumina), and cDNA libraries were purified using AMPure XP Beads (Beckman). Following end-repair and poly(A)-tailing, adaptor ligation was performed with Truseq RNA single indexes set A/B (Illumina).