GSM7911245: WT_clone_2h_r2; Mus musculus; ATAC-seq
50,000 macrophages were centrifuged and washed with PBS once. The cell pellet was resuspended with cold hypotonic lysis buffer. The lysate was centrifuged and the supernatant was discarded. The crude nuclei were treated with transposition reaction mix (50ul) containing 2.5ul Tn5 transposase (Nextera) and incubated in a 37oC thermomixer with 1000RPM mixing for 30 minutes. The transposed DNA was then cleaned up with a Qiagen Minelute PCR Purification Kit. The transposed DNA was amplified by a non-saturating PCR containing primer 1 and barcode primer 2 provided in the published protocols by Buenrostro et al. The amplified DNA was size selected on the gel from 100bp to 1000bp and purified with a Qiagen QIAquick Gel Extraction Kit. The selected DNA library was quantified by Qubit and Agilent Bioanalyzer High-Sensitivity DNA Analysis Kit.