GSM2231397: 5022-SUP_RRBS; Mus musculus; Bisulfite-Seq
The DNA methylation levels in Nr3c1 and Crebbp promoters in the hippocampi of F1m-GCS and F1m-CON mice were examined by DNA bisulfate conversion and PCR amplification and Sequencing using the EZ DNA Methylation-GoldTM Kit (Zymo Research, USA) following the manufacturer’s standard protocol. RRBS libraries for the hippocampi of F1m-GCS and F1m-CON were constructed as previously described(Cai et al., 2015; Gu et al., 2011). Briefly, 500 ng of genomic DNA was digested with 20 units of MspI (NEB, USA) in a 20 μl reaction for 20 h at 37 °C. After purification, the digested products were blunt-ended, and then dA was added, followed by methylated-adapter (Illumina, USA) ligation. To obtain DNA fractions in 40-120 bp and 120-220 bp range of MspI-digested products, two ranges (160-240 bp and 240-340 bp) of adapter-ligated fractions were excised from 2% agarose gel. The size-selected DNA was bisulfite-treated for two rounds using EpiTect Bisulfite Kit (Qiagen, German) following the manufacturer’s standard protocol. The final libraries were generated by amplification with HiFi HotStart Uracil + ReadyMix PCR kit (KAPA, USA). The quality and quantity were analyzed by an Agilent 2100 Bioanalyzer (Agilent Technologies, USA) and qPCR, respectively. Illumina HiSeq 2000 was used to sequence the libraries according to the manufacturer’s protocol.
GEO Accession
GSM2231397