Nuclei Isolation for Single Cell ATAC Sequencing

Demonstrated Protocol, For use with cell lines and PBMCs, Last Modified on July 8, 2019, Permalink

CG000169_DemonstratedProtocol_NucleiIsolation_ATAC_Sequencing_RevD.pdf

20190618_CG000169_Nuclei_Isolation_ATAC_Sequencing_RevCtoRevD.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.