GSM5962469: T293; Homo sapiens; ATAC-seq
The number of 293T cells and J76 cells in the logarithmic growth phase was counted by hemacytometer. 50000 cells were added to PCR tubes and centrifuged at 4 ºC at 600 x g for 5 min, discarded the supernatant. The pre-cooled 0.1% NP-40 lysis buffer solution was added to the cell precipitate and pipetted until homogeneous. The cells were lysed on ice for 3 minutes. After centrifugation at 4 ºCa t 550 x g for 5 min, discard the supernatant. The two kinds of DNA samples were cut by adding the self-made Tn5 transposase mixed system into the precipitation, and the library was constructed before ATAC-seq sequencing. The reaction system was10 µl 5 x TAPS-MgCl2-PEG 8000, 32 µl H2O, 8 µl 0.96 µg/µl Tn5 enzyme. The reaction conditions were the same as 1.2.3 enzyme activity detection.  The DNA of Tn5 reaction system samples was purified by an Omega PCR purification kit. The purified DNA samples were collected and amplified by PCR. The DNA samples were sent to Guangzhou Nuo He Zhi Yuan Company for sequencing. A portion of the DNA was used for Q-PCR to measure the concentration of DNA.