title: GSM2577161: E3.5_MethylC-Seq; Mus musculus; Bisulfite-Seq
library_construction_protocol: MethylC-Seq: Purified DNA spiked in with 0.1% unmethylated cl857 Sam7 lambda DNA (Promega) was sonicated using the microtube protocol-300 of Covaris S2 (Applied Biosystems). End-repair, dA-tailing and ligation were performed using the NEBNext End Repair Module, dA-Tailing Module and NEB T4 ligase. The adaptor-ligated DNA with insert size larger than 200 bp was selected by gel electrophoresis. Bisulfite conversion was performed using the EZ DNA methylation Gold Kit (Zymo Research) according to the manufacturer's instructions. The time of 64°C incubation was 2 hr for some libraries. Bisulfite- treated DNA was amplified using ZymoTaq DNA Polymerase or KAPA HiFi HotStart Uracil+ ReadyMix with 14-15 cycles.  Mouse gametes and embryos were collected at minimal volume and frozen. DNA was extracted using QIAamp DNA Mini Kit (QIAGEN) and RNA was isolated using RNeasy Micro Kit (QIAGEN).
attributes.tag: GEO Accession
attributes.value: GSM2577161