title: ATAC-seq of heterocephalus glaber: 4h_rep2
design: Library selection: A total of 50,000 cells were washed twice with 500 _l cold PBS and dissociated in 50 _l lysis buffer (10 mM Tris_HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% (v/v) Nonidet P-40 Substitute). The lysis reaction was terminated by adding a 20-fold large volume of ATAC-RSB reagent, and cell debris was removed. In order to obtain better quality nuclei, centrifuge at 4C and 500g at low speed. Nuclei were resuspended in 1x TAG buffer and stained with DAPI. Under the 10X objective of the upright microscope, the morphology of the nuclei was observed and the nuclei were counted. About 1105 nuclei were added to ATAC-transposase reaction reagent to capture the open area of chromatin by transposition. The transposition reaction was carried out at 37C for 30min. Transposase-acted DNA fragments were purified using MinElute PCR Purification Kit (QIAGEN, cat. #28004), and the purified products were subjected to PCR amplification, and ATAC library adapters were added during the PCR process. In order to determine the number of PCR cycles that need to be performed, the qPCR method was used to confirm the number of cycles through the amplification curve to reduce PCR redundancy. The ATAC library fragments recovered by magnetic bead purification were used to prepare and sequence the single-chain loop library using the BGISEQ500 sequencing platform, and the sequencing category was PE 50+32.