title: GSM8454255: Ctl1B_Day0_repl2; Homo sapiens; ATAC-seq
library_construction_protocol: ATAC-Seq libraries were prepared following. In brief, two technical replicates per sample containing 50,000 nuclei each were collected at 500 x g for 5 min at 4°C. Nuclei were resuspended in lysis buffer (10 mM Tris-Cl, pH 7.4; 10 mM NaCl; 3 mM MgCl2; 0.1% Igepal CA-630) and centrifuged at 500 x g for 10 min. The supernatant was removed and each nuclei pellet was resuspended in tagmentation buffer (25 μL 2 × TD buffer) with 2.5 μl Tn5 transposome and incubated at 37°C for 30 min. An equal volume of 2 x reverse crosslinking buffer (50 mM Tris-Cl, 1mM EDTA, 1% SDS, 0.2M NaCl, 5 ng/ml Proteinase K) was added to each sample and incubated at 65°C with agitation at 1200 rpm for overnight. DNA was isolated using a MinElute PCR Purification Kit (Qiagen). Sequencing libraries were prepared following described standard protocol. All libraries were sequenced on an Illumina Nova-seq platform at Novogene Europe. dsDNA was tagmented with Tn5, then PCR is performed using primers with barcode.