Demonstrated Protocol, For use with cell lines and PBMCs, Last Modified on September 9, 2020, Permalink
This protocol outlines how to isolate, wash, and count nuclei suspensions for use with the Chromium Next GEM Single Cell Multiome ATAC + Gene Expression (GEX) protocol (CG000338). Cryopreserved primary cells (PBMCs) and cell lines (GM12878 cells; 3T3 cells) were used to develop this protocol. PBMCs were cryopreserved in IMDM + 40% FBS + 15% DMSO. Cell lines were cryopreserved in RPMI + 15% FBS + 5% DMSO. Optimization of some protocol steps (e.g. lysis time, centrifugation speed/time and filtration steps) may be needed based on cell type.
This protocol is only compatible with the Chromium Single Cell Multiome ATAC + Gene Expression reagent kits. Do not use with standalone Single Cell ATAC or SC3'v3.1 assays.
FOR USE WITH:
Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Reagent Bundle, 16 rxns PN-1000283, includes:
Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Reagent Bundle, 4 rxns PN-1000285, includes:
Chromium Next GEM Chip J Single Cell Kit, 48 rxns PN-1000234
Chromium Next GEM Chip J Single Cell Kit, 16 rxns PN-1000230
Single Index Kit N Set A, 96 rxns PN-1000212
Dual Index Kit TT Set A, 96 rxns PN-1000215