Question: How can I optimize my nuclei suspension if I am using an untested tissue type in the Chromium Nuclei Isolation kit?
Answer: For untested tissue types, we recommend using a lysis timeline to optimize for your tissue type. For untested sample types, we recommend using the default workflow before performing any optimization. If the 10 minute lysis time is too harsh on your tissue and there are signs of over-lysis in the final nuclei suspension, we would recommend optimization by reducing the lysis time from 10 minutes (such as 8, 6, and 3 minutes). You can check the suspension under the microscope following lysis to determine the nuclei quality. There will be some additional loss during the filtration and clean-up steps, but checking the nuclei following lysis step can help determine the status of the prep (cells not completely lysed or nuclei overlysed). For tough tissue types (such as heart and muscle), mincing the tissue on a glass dish can increase surface area and improve the lysis process to recover more nuclei. If there is high debris, a 30µm strainer can be used at the end of the workflow to remove problematic large debris.
Products: Single Cell Gene Expression, Single Cell ATAC, Single Cell Multiome ATAC + GEX, Fixed RNA Profiling