title: Illumina HiSeq 2000 sequencing; ChIP-Seq of Gabpa in male C57Bl/6 mice liver
design: ChIP-Seq of Gabpa in male C57Bl/6 mice liver
library_construction_protocol: Male 12wk old C57Bl/6J mice were obtained from Charles River Laboratories (Brussels, Belgium) and maintained on a 12 h light-dark cycle with regular, unrestricted access to water. Pulverized liver tissue from 12 wks old male C57Bl/6J mice was fixed with 1 % formaldehyde in PBS extracts were homogenized by douncing in lysis buffer and chromatin fragmentation and ChIP assays were performed as described (Canettieri et al., 2003) using antibodies specific for GAbpα or unspecific IgG.Cross-linking was reversed overnight at 65°C, and DNA isolated using MinElute PCR Purification Kit. Enriched DNA from ChIP and Input DNA fragments were end-repaired, extended with an 'A' base on the 3end, ligated with indexed paired-end adaptors using the Bravo Platform, size-selected after 4 cycles of PCR with AMPure XP beads and amplified by PCR for 10 cycles more. Libraries were prepared from approximately 50 pg of ChIP DNA using the ThruPlex-FD Prep Kit