title: GSM1471901: RRBS E8.5_WT-Rep1; Mus musculus; Bisulfite-Seq
library_construction_protocol: Genomic DNA was extracted by proteinase K digestion followed by phenol/chloroform extraction. We prepared RRBS libraries using a published protocol (Gu et al., Nat Protoc 6:468-81, 2011) with some modifications. Briefly, we digested 25-100 ng of genomic DNA 5 hours with MspI (Fermentas), followed by end-repair and A-tailing (Klenow fragment, Fermentas), and ligation to methylated adapters (T4 DNA ligase, Fermentas) in Tango 1X buffer. Fragments between 150 and 400 bp were excised from a 3% agarose 0.5X TBE gel with the MinElute kit (Qiagen) and bisulfite-converted with the EpiTect Bisulfite kit (Qiagen) using two consecutive rounds of conversion. Final RRBS libraries were amplified with 14-16 cycles of PCR.