title: GSM1173801: hmC-gluc-IP-Seq_mm_fc_6wk; Mus musculus; OTHER
library_construction_protocol: Genomic DNA was purified from cells using the DNeasy Blood and Tissue Kit (Qiagen, Valencia, CA, USA) according to the manufacturer's instructions. hmC-gluc-IP-Seq sequenging libraries were generated by biotin-glucosyl tagging and enrichment of hmC according to the following procedure. The Hydroxymethyl Collector Kit (Active Motif, Carlsbad, CA, USA) was used to selectively tag hmC bases within genomic DNA fragments (100-200 bp) with glucose and biotin moieties to form biotin-N3-5-gmC (biotin-gmC), as described previously [Song C.-X. et al., Selective chemical labeling reveals the genome-wide distribution of 5-hydroxymethylcytosine, Nature Biotechnology 29, 68–72 (2011)]. Genomic DNA fragments containing biotin-gmC were then enriched and purified by high-affinity capture using streptavidin magnetic beads. Enriched DNA containing biotin-gmC was used to generate Illumina DNA sequencing libraries with the TruSeq DNA Sample Prep Kit v2 (Illumina, San Diego, CA).