GSM8391376: GFI1KO, scATAC; Mus musculus; ATAC-seq
Cells were pelleted for 5 mins at 300 × g followed by nuclei isolation by incubating cells with chilled cell lysis buffer (10 mM Tris-HCl, 10 mM NaCl, 3 mM MgCl2, 0.1% tween-20, 0.1% IGEPAL CA-630, 0.01% digitonin, 1mM dithiothreitol, 1 U/μL RNase inhibitor and 1% BSA) for 3 min. Nuclei were washed twice in 1 ml of wash buffer (10 mM Tris-HCl (pH 7.4), 10 mM NaCl, 3 mM MgCl2, 0.1% Tween 20, 1 mM dithiothreitol and 1 U/µL RNase inhibitor and 1% BSA) by centrifuging at 500 × g for 5 min. After centrifugation, cells were resuspended in chilled Nuclei Buffer (1× Nuclei Buffer, 1 mM dithiothreitol and 1 U/µL RNase inhibitor). Nuclei were incubated in transposition mix according to the Chromium Next GEM Single Cell Multiome ATAC + GEX user guide (protocol CG000338 Rev F). Following transposition, GEMs were generated using 10x Chip J. Sample cleanup and amplification PCR were performed as per the user guide.