Nuclei Isolation for Single Cell ATAC + Gene Expression Sequencing

Demonstrated Protocol, For use with cell lines and PBMCs, Last Modified on September 9, 2020, Permalink

CG000365_DemonstratedProtocol_NucleiIsolation_ATAC_GEX_Sequencing_RevA.pdf

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: