title: GSM3731617: HuES64_HIC2699_MboI; Homo sapiens; Hi-C
library_construction_protocol: Hi-C libraries were prepared following the protocol described in Rao et. al. 20141. Briefly, one million cells were crosslinked with 1% formaldehyde for 10  minutes at room temperature and then quenched with 0.2M glycine solution. Cells were lysed and nuclei permeabilized with 0.5% sodium dodecyl sulphate for 10 minutes at 62°C. Chromatin was digested with 100 U of MboI restriction enzyme (New England Biolabs). Ends of the restriction fragments were blunted and labeled with a biotinylated nucleotide and then ligated. Nuclei were pelleted, proteins were digested with proteinase K and crosslinks were reversed by heating at 68°C overnight. DNA was sheared in a Covaris focused ultrasonicator to average fragment length of 400 bp. Size-selected DNA was enriched for biotinylated ligation products through binding to T1 streptavidin beads (Thermo Fisher). Libraries were prepared for Illumina sequencing by performing the end-repair, A-tailing and adapter ligation steps with DNA attached to the beads. Hi-C libraries were amplified directly off the beads and purified for subsequent Illumina sequencing with 100 paired-ends.
attributes.tag: GEO Accession
attributes.value: GSM3731617