GSM2837155: R836A_eRRBS, biological replicate 3; Mus musculus; Bisulfite-Seq
Genomic DNA was extracted using DNeasy Blood & Tissue Kit (Qiagen). eRRBS was carried out with a previously described protocol with slight modification (Garrett-Bakelman et al., 2015). Briefly, ~300 ng of genomic DNA were digested with three different enzymes (~80 units of MspI, 40 units of BfaI and 40 units of MseI) to enhance genomic fragmentation and coverage. The produced fragments were ligated to pre-annealed adapters containing 5'-methyl-cytosine instead of cytosine, followed by overhang fill-in, 3'-terminal-A extension and purification. Bisulfite treatment of the fragments was done using the EZ DNA Methylation–Lightning kit (Zymo Research). Amplified eRRBS libraries was quality checked with Agilent 2200 TapeStation, followed by deep sequencing on the Illumina HiSeq-2000 genome analyzer with 50 bp PE parameters.
GEO Accession
GSM2837155