Question: Are there differences in the Chromium Nuclei Isolation protocol when isolating nuclei for Single Cell Gene Expression applications, Single Cell Multiome ATAC + GEX, and Single Cell ATAC?
Answer: There are a few important points when considering Isolating nuclei for the various compatible downstream applications.
Single Cell Gene Expression (3’/5’/Fixed RNA) |
Single Cell Multiome ATAC + Gene Expression |
Single Cell ATAC |
|
Kit used |
Chromium Nuclei Isolation Kit with RNase Inhibitor, 16rxns |
Chromium Nuclei Isolation Kit with RNase Inhibitor, 16rxns |
Chromium Nuclei Isolation Kit, 16rxns |
Lysis Buffer |
Generate Lysis buffer as instructed |
Generate Lysis buffer as instructed |
Generate lysis buffer as instructed, additional dilution with 1x PBS |
RNase Inhibitor |
Wash and resuspension buffers include RNase Inhibitor |
Wash and resuspension buffers include RNase Inhibitor |
Wash and resuspension buffers do not include RNase Inhibitor |
Final nuclei resuspension |
Use wash and resuspension buffer |
Use 1x Nuclei Buffer with RNAse inhibitor |
Use 1x Nuclei Buffer |
Products: Single Cell Gene Expression, Single Cell Immune Profiling, Single Cell ATAC, Single Cell Multiome ATAC + GEX, Fixed RNA Profiling