GSM5219357: OSKM_D9_R1_R2 (RRBS); Mus musculus; Bisulfite-Seq
The RRBS libraries were sequenced by Illumina HiSeq 2000 platform. RRBS assay was performed as previously described (Boyle et al., 2012), briefly, 20ng of genomic DNA were digested with Msp1 restriction enzyme (NEB, R0106L), DNA fragments were end-repaired and A-Tailed using Klenow fragment (3'-5- exo-) (NEB, M0212L), the DNA fragments were then size selected using AMPure XP beads (Beckman Coulter Genomics, A63881) and ligated to illumina adaptors (Illumina, PE-940-2001) using T4 ligase (NEB, M0202M). The samples were then subjected to two consecutive bisulfite conversions using EpiTect Bisulfite Kit (QIAGEN, 59104) and PCR using PfuTurbo Cx hotstart DNA polymerase (Agilent Technologies, 600412).