alias: GSM2465666
title: GSM2465666: Heart_1wk_M04NB; Mus musculus; Bisulfite-Seq
library_construction_protocol: All tissues were snap frozen directly after isolation. Genomic DNA was isolated from ~10mg frozen tissue using the DNeasy Blood & Tissue Kit (Qiagen). RRBS libraries were prepared from isolated DNA following published protocols (Smallwood et al.,2011). Briefly, RRBS libraries were prepared by MspI digestion of 100-500ng genomic DNA, followed by end-repair and T-tailing using Klenow Exo- (Fermentas). Adapter ligation was performed overnight (homemade adapters) using T4 DNA Ligase (NEB), followed by a cleanup step using AMPure XP beads (Agencourt, 0.9x). Subsequently, libraries were bisulfite treated according to the manufactures instructions (Sigma Imprint Kit; 2 step protocol) and purified using an automated liquid handling robotic system (Agilent Bravo). The libraries were amplified using KAPA HiFi Uracil HotStart DNA Polymerase (KAPA Biosystems), indexing the samples with individual primers. All amplified libraries were purified (AMPure XP beads, 0.8x) and assessed for quality and quantity using High-Sensitivity DNA chips on the Agilent Bioanalyzer. High-throughput sequencing of all libraries was carried out with a 75 bp paired-end protocol on a HiSeq 2000 instrument (Illumina).
identifiers.namespace: GEO
identifiers.id: GSM2465666