Question: Do I need to sort to remove debris after isolating nuclei using the Chromium Nuclei Isolation kit?
Answer:
- No additional sorting has been required for any samples tested post nuclei isolation.
- Debris removal steps in the Chromium Nuclei Isolation kit workflow remove large and small debris from debris-heavy samples (i.e., brain).
- In the example below, a myelin debris-heavy adult mouse brain was dissociated using the 10x Genomics Chromium Nuclei Isolation kit (bottom left). The sample was taken through the entirety of the workflow resulting in a clean nuclei suspension (top right). When the same dissociated sample is passed through a 70 um filter (bottom right), lots of large and small debris remains within the sample.
- Some small debris in the final nuclei suspension is acceptable and expected for debris-heavy sample types and will not negatively impact the assay.
- If an unacceptable amount of debris is still present after running through the workflow, an extra straining step (30µm strainer) can be added to further reduce debris.
Products: Single Cell Gene Expression, Single Cell Immune Profiling, Single Cell ATAC, Single Cell Multiome ATAC + GEX, Fixed RNA Profiling