title: GSM8239658: A5_ATAC-seq_2; Homo sapiens; ATAC-seq
library_name: GSM8239658
library_construction_protocol: Assay for transposase accessible chromatin was performed on hiPSC-derived hNPC using a derived protocol from Buenostro et al., 2013. Indeed, 50 000 cells from 8330 (control line CHD1L+/+), A12 and A5 (mutant for CHD1L) lines were resuspended in resuspension buffer (10mM Tris-HCl pH 7.5, 10mM NaCl, 3mM MgCl2) and then lysed in lysis buffer (Resuspension buffer + 0.1% NP40 + 0.1% tween-20+ 0.01% Digitonin) and incubated 3mn on ice. Then, 1mL of wash buffer was added (resuspension buffer + 0.1% Tween-20). Cells were centrifugated 10mn at 500G at 4°C and supernatant was trashed out. Transposition reaction mix (Kit Illumina Tagment DNA Enzyme and Buffer Small Kit #20034197) was added to pellet and incubated 30mn at 37°C in a thermomixer at 1000rpm. DNA fragment was isolated using Qiagen MinElut Reaction Cleanup kit. DNA fragment were amplificated and library was generated by PCR using appropriate primers as described in Buenostro et al., 2013 and NEBNext High-Fidelity 2X PCR Master mix (NEB, #M0541S). Finally, library was purified using SRIselect beads (Beckman Coulter) by a one-sided purification to remove primers.