GSM4308592: Cdk6 knock out (KO3) [RRBS]; Mus musculus; Bisulfite-Seq
Genomic DNA was extracted using Qiagen DNeasy kit. Genomic DNA was MspI (NEB) digested, end-repaired and A-tailed using Klenow Polymerase (NEB) followed by adapter ligation using Quick Ligase (NEB) and AMPure XP size selection (Beckman Coulter, Fullerton, CA, USA). RRBS libraries were sodium bisulfite treated using the EZ-DNA Methylation-Direct kit (Zymo Research Corp, Orange, CA, USA) and quantified by qPCR. Enrichment PCR was performed using the PfuTurboCx Hotstart Kit (Agilent Technologies, Santa Clara, CA, USA) followed by AMPure XP clean up. Quality of final RRBS libraries was determined by Experion analysis (Biorad). Sequencing was performed on a HiSeq2000 sequencer (Illumina Inc, San Diego, CA, USA).