GSM4257634: T3_ATAC-seq; Homo sapiens; ATAC-seq
a total of 50,000 cells were washed once with 50 mL of cold PBS and resuspended in 50 mL lysis buffer (10mMTris-HCl pH 7.4, 10mMNaCl, 3 mMMgCl2, 0.2% (v/v) IGEPAL CA-630). The suspension of nuclei was then centrifuged for 10 min at 500 g at 4 oC, followed by the addition of 50 mL transposition reaction mix (25 mL TD buffer, 2.5 mL Tn5 transposase and 22.5 mL nuclease-free H2O) of Nextera DNA library Preparation Kit (96 samples) (FC-121-1031, Illumina, USA). Samples were then PCR amplified and incubated at 37oC for 30 min. DNA was isolated using a MinElute Kit (QIAGEN, Germany). ATAC-seq libraries were first subjected to 5 cycles of pre-amplification. To determine the suitable number of cycles required for the second round of PCR the library was assessed by quantitative PCR as described (Buenrostro et al., 2015a), and the library was then PCR amplified for the appropriate number of cycles. Libraries were purified with a Qiaquick PCR (QIAGEN, Germany) column. Library concentration was measured using a KAPA Library Quantification kit (KK4824) according to the manufacturer's instructions
GEO
GSM4257634