GSM5671140: female KO rep2 [MUP12]; Mus musculus; Bisulfite-Seq
At twelve weeks of age mice were sacrificed by cardiac puncture and liver samples were taken and snap frozen. Genomic DNA was extracted using the All Prep DNA/RNA Extraction Kit (QIAGEN). The gDNA was digested with  MspI (Tango Buffer.Thermo Scientific), followed by nucleotide end-repair and A-tailing (Klenow Fragment exo- with dCTPs, dGTPs, and dATPs in Tango Buffer, Thermo Scientific) and heat inactivation.  EM-seq Adaptors (NEB E7165A, New England Biolaps) were ligated to the A-tailed DNA fragments using T4 DNA ligase (HC; Thermo Scientific). Bisulfite conversion was performed using ZYMO EZ-96 DNA Methylation-DirectTM MagPrep Kit.  Bisulfite converted libraries were amplified  Kapa (Uracil+ 2x MasterMix ;Roche) and purified (0.8x SPRIselect; Agencourt, Beckman Coulter).