GSM1648238: S25NC; Mus musculus; Bisulfite-Seq
We isolated genomic DNA from flash frozen livers using a phenol-chloroform extraction We digested 1μg of DNA with MspI restriction enzyme (NEB, Ipswich, MA, USA), carried out end-repair/adenylation (NEB) and ligation with TruSeq barcoded adapters (Illumina, San Diego, CA, USA). We selected DNA fragments of size range 200-300bp with AMPure magnetic beads (Beckman Coulter, Brea, CA, USA), followed by bisulfite treatment on the DNA (Millipore, Billerica, MA, USA), and PCR amplification (Bioline, Taunton, MA, USA ). We sequenced the libraries by multiplexing two libraries per lane in an Illumina HiSeq sequencer, with 100bp reads.