GSM2533297: ES_5hMeDIP_Dnmt3aKO; Mus musculus; OTHER
Genomic DNA was sonicated to 150-300 bp and RNAseA treated prior to end-repair and A-tailing using the NEB Ultra library kit and protocol. NEB adapters were ligated to the genomic DNA following the NEB Ultra protocol, and DNA was cleaned up using Ampure XP beads. Following cleanup, 5 ug adapter-ligated DNA was taken up in 500 ul TE and denatured for 10 min in boiling waterbath. 5-hmC was enriched using the Active Motif 5-hmC antibody (39769) following the MeDIP protocol from Mohn et al 2009, PMID:18987806. After IP, library was PCR amplified with 15-18 cycles prior to sequencing.