GSM4975207: NaiveB_RRBS_rep2; Mus musculus; Bisulfite-Seq
Genomic DNA was isolated from B1-8hi splenic B cells using chloroform extraction and ethanol precipitation. RRBS libraries were generated using 20 ng genomic DNA, subjected to MspI digestion (NEB), end repair/dA-tailing reaction using Klenow 3'-5' exo- (NEB) and ligation by Illumina sequencing adapters using T4 DNA ligase (NEB). The 150-350 bp fragments was extracted from gel, and treated with sodium bisulphite using EZ DNA Methylation-Gold Kit (Zymo Research). Followed by the library amplification and indexing using KAPA HiFi Hot-Start Uracil+ ReadyMix (Kapa Biosystems), purified libraries were sequenced on HiSeq 2500 platform (Illumina). The methylation status of each cytosine was calculated using the Bismark methylation.