GSM8958372: SNCAiso_rep3; Homo sapiens; ATAC-seq
Approximately 1 × 1,000,000 iPSC-derived neurons were resuspended in 47.5 µl lysis buffer (10mM Tris-HCl pH 7.5, 10 mM NaCl, 3 mM MgCl2, 0.1% IGEPA, CA-630 in water) at RT. After the addition of 2.5 µl DNA enzyme I (Illumina), cells were incubated at 37°C for 30 min. DNA was purified with Zymo ChIP DNA concentrator columns (Zymo Research), eluted with 11 µl of elution buffer, and amplified using NEBNext High-fidelity 2× PCR MasterMix (New England BioLabs) with the Nextera primer Ad1 (1.25µM) and a unique Ad2.n barcoding primer (1.25µM) for 8-10 cycles. The resulting libraries were size selected by gel excision to 165-250 bp, purified, and single end sequenced using a NovaSeq 6000 (Illumina) sequencer for 51 cycles according to the manufacturer's instructions.