Ensuring Quality Cell Input for Single Cell Sequencing Experiments by Viability and Singlet Enrichment Using Cell...

As with every sensitive analysis technology, the golden principle of "input quality rules output quality" also applies to single cell sequencing methods. Given the sensitivity of the current methods in single cell sequencing and the minuscule amounts of RNA present within a single cell, any extrinsic source of variability should be reduced by ensuring a homogenous input right at the start. Not every tissue is as readily handled as a single cell suspension like blood and most tissues will have to undergo digestions to free the cells from their spatial organization to undergo single cell transcriptomics workflows. This chapter provides working protocols for two simple, but very precise and powerful methods to ensure only the most viable cells are introduced into single cell assays.