GSM2412498: Ex.1 HDF TWIST1 locus; Homo sapiens; OTHER
Chromosome conformation capture (3C) was performed as described previously (Lieberman-Aiden et al., 2009) in HDF and iPSC. The restriction enzyme HindIII was used for digesting DNA in the crosslinked nuclear. 3C library was purified by phenol chloroform extraction and PureLink® Genomic DNA Mini Kit (Invitrogen). 3C libraries were sonicated into 300 to 700 bp by covaris E210 system. Ends of the sonicated 3C library were repaired and A-tailed by NEBNext® Ultra™ End Repair/dA-Tailing Module (NEB E7442S) according to manufacturer's instructions. Splinkerette adapters were ligated to the dA-tailed 3C libraries by Ligation high Ver.2 (TOYOBO LGK-201). Adaptor ligated 3C libraries were purified with 1.8 x AMPureXP beads. Target (bait) loci were enriched by nested PCR using universal primers and bait primers. The amplified 3C libraries were resolved on 2% agalose gel, and the libraries between 350 bp and 700 bp were excised and purified with QIAquick Gel Extraction Kit (QIAGEN 28704). Quantification of the ms3C-Seq libraries was performed by Library Quantification Kits (KAPA KK4824)