ATAC-seq of sorted cells at day 7.5 of cardiac differentiation from human embryonic stem cells
ATAC-seq of sorted cells at day 7.5 of cardiac differentiation from human embryonic stem cells
Embryoid bodies on differentiation day 7.5 were dissociated to single cells by TrypLE Select and purified by FACS based on NKX2-5-GFP and SOX17-Tomato reporter status and immediately processed for ATAC-seq library preparation. ATAC-seq libraries were prepared using an ATAC-Seq Kit (Active Motif) as per the manufacturer's instructions. Briefly, cells were gently washed with ice cold PBS and lysed with ATAC lysis buffer. Tagmentation master mix was prepared (Tagmentation buffer, PBS, 0.5% Digitonin, 10% Tween 20, Assembled Transposomes) and tagmentation carried out at 37oC for 30 minutes at 800 rpm. DNA was purified and then PCR amplified using unique combinations of i7 and i5 indexing primers per sample. After PCR amplification, samples were purified using SPRI beads and quality determined using Agilent Tapestation 2200. ATAC-seq libraries were prepared using an ATAC-Seq Kit (Active Motif) as per the manufacturer's instructions.