title: GWAS region focused capture-HiC of primary melanocytes from five human donors
design: GWAS region focused capture-HiC of primary melanocytes from five human donors
library_construction_protocol: Early passages of primary human melanocytes were isolated from foreskin healthy newborn males, following an established protocol (Halaban et al, J Exp Med (2000) 191(6):1005-1016) from the Specialized Programs of Research Excellence (SPORE) in Skin Cancer Specimen Resource Core at Yale University. Cells were grown in Dermal Cell Basal Medium (American Type Culture Collection/ATCC PCS-200-030) supplemented with Melanocyte Growth Kit (ATCC PCS-200-041) and 1% amphotericin B/penicillin/streptomycin (120-096-711, Quality Biological). C56, C140 and C205 are of European ancestry(CEU) and C24 and C27 are of African descent(YRI). For each library, 50,000 live cells were harvested. The initial Hi-C libraries were generated using the Arima HiC kit (Arima Genomics) following the manufacturer's protocol. Briefly, 2-4 million cells were crosslinked, enzyme digested, and ligated. The ligated DNA were purified by Ampure beads. The ligated DNA was reverse-crosslinked, fragmented by sonication, and size-selected (200-600bp) for adaptor ligation and library amplification using KAPA HyperPrep kit (KAPA Biosystems) following the manufacturer's protocol.The HiC library was then hybridized with the custom capture baits and captured by the SureSelect XT HS and XT low input library preparation kit for ILM (Agilent).