title: Assay for Transposase-Accessible Chromatin sequencing (ATAC-seq) of primary melanocytes from five human donors
design: Assay for Transposase-Accessible Chromatin sequencing (ATAC-seq) 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 ATAC-seq libraries were generated following a previously published protocol (Buenrostro et al., Nature Methods, 2013). The nuclei were isolated from the 50,000 cells. The isolated nuclei were treated with Tn5 Transposes from Nextera DNA Sample Preparation Kit and the transposed DNA was purified using a Qiagen MinElute Kit. The transposed DNA libraries were amplified by PCR for 12 cycles and isolated by Ampure XP beads.