GSM5743950: MTP _rep1A; Homo sapiens; ATAC-seq
Library for ATAC-seq was prepared according to a published protocol (Buenrostro J D.et al., 2013) using 50,000 intact cells as input. The cells were spun at 500g for 5 min followed by a wash using 50 μL of cold 1× PBS and centrifugation at 500g for 5 min. Cells were lysed using cold lysis buffer (10 mM Tris-HCl, pH 7.4, 10 mM NaCl, 3 mM MgCl2 and 0.1% IGEPAL CA-630) and spun at 500g for 10 min using a refrigerated centrifuge generating the nuclei prep. The nuclei prep was then resuspended in the transposase reaction mix (25 μL 2× TD buffer, 2.5 μL transposase (Illumina) and 22.5 μL nuclease-free water) 30 min at 37 °C. Tn5 transposase tagmentation simultaneously fragments the genome and tags the resulting DNA with Illumina sequencing adapters. DNA fragments were PCR ampliﬁed and subsequently puriﬁed using Qiagen MinElute PCR Puriﬁcation Kit (Qiagen, Hilgen, Germany). Final library quality and quantity were analyzed and measured by Agilent Bioanalyzer 2100 (Agilent Technologies, Santa Clara,CA, USA) and Life Technologies Qubit3.0 Fluorometer (Life Technologies), respectively.