GSM6519819: direct Neurons D868N ATAC #1; Homo sapiens; ATAC-seq
ATAC-seq was performed using a previously described protocol 3. Briefly, 50000 of cells for each experimental replicated were resuspended in lysis buffer (10 mM Tris-HCl, pH 7.4, 10 mM NaCl, 3 mM MgCl2 and 0.1% IGEPAL CA-630, Sigma aldrich) to perform crude nuclei isolation. Trasposition reaction was then performed using Tn5 transposase (Illumina).  After transposition, DNA was purified using Zymo DNA Clean & Concentrator kit (cat. D4013) and amplified by PCR (NebNext ultra II polymerase, cat. M0544S). After 5 cycles the reaction was stopped, the necessary amount of cycles needed for the final amplification were calculated with qPCR. After amplification libraires were purified using Ampure XPbeads selection fragments between 100-1000 bp. Subsequently samples were sequenced paired-end on NovaSeq6000 by GeneWiz. NexTera