GSM8831243: Pooled vs Parallel - Parallel ATAC; Homo sapiens; ATAC-seq
Cell suspensions were first washed once with chilled PBS. 500k cells per sample were aliquoted into 1.5mL Eppendorf tubes and pelleted at 300rcf, 4˚C, for 4 minutes. Cells were resuspended in 100 µL of chilled Lysis Buffer (10 mM Tris-HCl pH 7.4, 10mM NaCl, 3mM MgCl2, 0.1% Tween-20, 0.1% Nonidet P40 Substitute, 0.01% Digitonin, 2% BSA in nuclease-free water), mixed, and incubated 5 minutes on ice. Then, 1 mL Wash Buffer (10 mM Tris-HCl pH 7.4, 10mM NaCl, 3mM MgCl2, 0.1% Tween-20, 2% BSA in nuclease-free water) was added and mixed. Nuclei were pelleted at 500rcf, 4˚C, for 4 minutes and then resuspended in chilled PBS. See MULTI-ATAC manuscript for more details. Nuclei were transposed and subsequently processed into scATAC-seq or Multiome libraries according to manufacturer's recommendations (10x Genomics), with only minor modifications. A 1 µL aliquot is taken from each individual library to be used in producing accompanying MULTI-ATAC barcode libraries. See MULTI-ATAC manuscript and protocol for more details.