Question: How do I dissociate my tissue of interest?
Answer: We have not tested dissociating all tissue/sample types. General guidelines for preparing optimal single cell suspensions can be found in the Cell Preparation Guide (https://support.10xgenomics.com/single-cell-gene-expression/index/doc/demonstrated-protocol-single-cell-protocols-cell-preparation-guide).
Below are some resources that may be helpful when considering methods for dissociating your tissue of interest.
1. Demonstrated Protocol - Dissociation of Mouse Embryonic Neural Tissue for Single Cell RNA Sequencing (https://support.10xgenomics.com/single-cell-gene-expression/index/doc/demonstrated-protocol-dissociation-of-mouse-embryonic-neural-tissue-for-single-cell-rna-sequencing)
Although written for mouse embryonic neural tissue, this protocol provides a good overview on how to prepare fresh tissue for the Single Cell 3' assay. The specific enzymes and incubation times may be different depending on tissue type and will need to be optimized. Generally, the cell viability should be > 80% and and between ~200 and 18,000 cells should be loaded per sample. If this is the first time a specific cell/tissue type is being run, it is generally recommend to target ~2,000 cells.
2. Commercially available solutions for tissue dissociation
Several companies have products designed specifically for tissue dissociation; we recommend consulting their websites for information: i) Miltenyi Biotec, ii) Worthington Biochemical, iii) Sigma-Aldrich, and iv) Roche Diagnostics. Some of the products have been validated in house (e.g. Miltenyi Micro Beads).
3. Published dissociation methods
We recommend the Worthington Biochemical website for recommendations and published protocols on specific tissue types: http://www.worthington-biochem.com/tissuedissociation/default.html
Products: Single Cell 3', VDJ