GSM2060311: GLIB_5hmC.Tet2.KO; Mus musculus; OTHER
Chemical labeling-based 5hmC enrichment was performed according to Hydroxymethyl Collector™ kit from Active Motif (cat#55013, Active Motif-Carlsbad, CA) to selectively label 5hmC bases with glucose and biotin moieties to form biotin-N3-5-gmC as previously described by Song et al (Song CX, et al. (2011) Selective chemical labeling reveals the genome-wide distribution of 5-hydroxymethylcytosine. Nature biotechnology 29(1):68-72). Briefly 4 ug of genomic DNA was sonicated with COVARIS S2 to target size 200-400 bp (250 bp protocol for 150 seconds), purified with 1X AMPure XP beads (Agencourt A63880) and then subjected to biotin-based enrichment of 5-hydroxymethylcytosine following the recommended procedure. Captured 5hmc DNA enriched fragments were then processed for Illumina sequencing as previously described (Garber et al. 2012) with slight modifications (Ostuni et al. 2013).
GEO Accession
GSM2060311