GSM4626796: Tr4_2; Mus musculus; ATAC-seq
The cell preparation was begin with harvesting and counting intact and homogenous cell, which followed by centrifuging those 50,000 cells for about 5min at 500g, 4°C. Washing those cells once with 50μl of cold PBS buffer and centrifuging them again under the same condition after removing and discarding supernatant. Removing and discarding supernatant again， and pipetted up and down to resuspend the cell pellet in 50μl of cold lysis buffer gently, which also obey centrifuge but for 10 min this time. After discarding the supernatant, the resuspension of the nuclei pellet we obtained should be carried out immediately in the prepared transposition reaction mix which contained Nextera Tn5 Transposase from Nextera kit. Incubated the transposition reaction at 37°C for 30 min. A Qiagen MinElute PCR Purification Kit is used to transposition, purify instantly, following by which the 10μl elution buffer (Buffer EB from the MinElute kit consisting of 10 mM Tris-HCl [pH 8.0]) was applied to elute transposed DNA. Finally the PCR amplification was performed to amplify transposed DNA fragments, combine the following in a 0.2 ml PCR tube with enough thermal cycle. As is known to us, prior to amplification, adapters have to be completed with a 72°C extension step, as well as the qPCR and library quality control using gel electrophoresis is provided. Libraries were pooled at equimolar ratios with barcodes and sequenced on the BGISEQ-500 platform (BGI-Shenzhen, China)
GEO Accession
GSM4626796