title: GSM1873375: WGBS_ESC_KO; Mus musculus; Bisulfite-Seq
library_construction_protocol: DNA was purified using QIAQuick PCR Purification Kit (QIAGEN) according to the manufacture’s instruction. Total RNA was extracted by using TRIzol reagent (Invitrogen). For whole-genome Bisulfite-Seq library preparation, 2.5 micrograms of ESCs genomic DNA, were spiked-in with 1ng of Escherichia coli genomic DNA, and sheared using a Bioruptor® Twin sonicator (Diagenode) for 3 runs of 10 cycles [30 s “ON,” 30 s “OFF”] at high power setting. Fragmented/digested DNA was then end-repaired, dA-tailed, and ligated to methylated adapters, using the Illumina TruSeq DNA Sample Prep Kit, following manufacturer instructions. DNA was loaded on an EGel Size select 2% agarose pre-cast gel (Invitrogen), and a fraction corresponding to fragments ranging from 180bp to 350bp was recovered. Purified DNA was then subjected to bisulfite conversion using the EpiTect Bisulfite Kit (Qiagen). Bisulfite-converted DNA was finally enriched by 15 cycles of PCR using Pfu Turbo Cx HotStart Taq (Agilent).
attributes.tag: GEO Accession
attributes.value: GSM1873375