Liver_Input
3 DNA pools of 2 mouse livers in each, the lean and obese groups were subjected to 5meDIP-seq. After extraction and sonication of DNA, Illumina sequencing adaptors were ligated to the DNA ends. Methylated DNA (meDNA) was immunoprecipitated using the 33D3 antibody (Active Motif, Carlsbad, CA, USA). Specific pulldown of 5meC DNA was confirmed by real-time PCR using primers to known methylated sites in the Zc3h13 and Grik3 genes and to one unmethylated site in the Untr6 gene. Next generation sequencing libraries were generated from the DNA immunoprecipitates and non-immunoprecipitated input DNA pooled from all 6 mouse livers and subjected to single-end sequencing on Illumina's Hi-Seq 2000 sequencer.