This protocol outlines how to isolate, wash, and count single nuclei from fresh, cryopreserved, and flash frozen mouse brain tissue samples for use with the Chromium Single Cell ATAC Solution. Tissue triturated into a nearly single cell suspension and subsequently frozen in media containing 10% DMSO produced metrics comparable to fresh tissue. High quality data can also be obtained using flash frozen tissue. Optimization of some protocol steps (e.g. homogenization, lysis reagent/time, centrifugation speed/time and filtration steps) may be needed when working with mouse brain tissues from different sources.
FOR USE WITH:
- Chromium Single Cell ATAC Reagent Kits User Guide (v1.1 Chemistry) (CG000209)
- Chromium Single Cell ATAC Reagent Kits User Guide (v1 Chemistry) (CG000168)
- Chromium Next GEM Single Cell ATAC Reagent Kits v2 User Guide (CG000496)