title: Illumina HiSeq 2000 paired end sequencing; Transcription profiling by high throughput sequencing of pancreatic islets from R299Q AMPK gamma 2 knock-in mice
design: Transcription profiling by high throughput sequencing of pancreatic islets from R299Q AMPK gamma 2 knock-in mice
library_name: WT_5
library_construction_protocol: Pancreatic islets (50-200) were extracted from  homozygous and heterozygous R299Q AMPK g2 knock-in mice and their wild type littermate controls aged 11-13 weeks by pancreatic distension and digestion with collagenase. Islets were incubated in RPMI media containing 11 mM glucose, 10% FCS, 100 IU/mL penicillin, 100 Âľg/mL streptomycin, at 5% CO2 and 37 C for 24 h. RNA was extracted with TRIzol (Invitrogen) using the manufacturer's protocol, with the addition of glycogen as a co-precipitate. Resuspended RNA was treated with DNase I (Invitrogen), which was then heat inactivated. Polyadenylated RNA was purified with oligo-dT beads and the library constructed using the TruSeq Stranded kit (Illumina). Illumina Universal Paired-end adapters were used, and custom barcoding sequences incorporated during the amplification rounds (15 cycles) for multiplexing.
attributes.tag: Experimental Factor: wild type genotype
attributes.value: genotype