Tumor Dissociation for Single Cell RNA Sequencing

Demonstrated Protocol, Last Modified on March 31, 2021, Permalink

CG000147_Demonstrated_Protocol_Tumor_Dissociation_RevB.pdf

This Demonstrated Protocol outlines best practices for dissociation of mouse melanoma, colon tumor, and breast tumor for use in 10x Genomics® Single Cell protocols.

While this protocol is demonstrated with freshly obtained (not frozen) mouse melanoma, colon tumor, and breast tumor, it may be used for dissociating other tumor types for use in 10x Genomics Single Cell protocols. Modifications to this demonstrated protocol may be necessary for other tumor types (e.g. dissociation time, resuspension buffer, enzyme concentration, centrifugation speed and time).

This demonstrated protocol is optimized for counting cells in the range of 700-1200 cells/µl. If using the Single Cell 3' LT v3.1 (low throughput) application, ensure cells are counted as indicated in this protocol and then diluted to the LT specific optimal loading concentration of 100-600 cells/µl using the Cell Dilution Overview in the LT User Guide.