Nuclei Isolation for Single Cell ATAC Sequencing

Demonstrated Protocol, For use with cell lines and PBMCs, Last Modified on March 24, 2022, Permalink

CG000169_DemonstratedProtocol_NucleiIsolation_ATAC_Sequencing_Rev E.pdf

This protocol outlines how to isolate, wash, and count nuclei suspensions for use with the Chromium Single Cell ATAC protocol. Cryopreserved primary cells (PBMCs) and cell lines (GM12878 cells; EL4 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 ATAC reagent kits. Do not use with Multiome ATAC + Gene Expression or SC3'v3.1 assays.