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1 1 This protocol is designed to obtain blastema cells (and wound epidermis if left attached) into a single cell suspension. It was specifically designed for single cell RNA sequencing, but can be used for other applications in which a single cell suspension is desired. The goal is to obtain a single cell suspension with near 100% viability in a rapid fashion. This protocol does not use centrifugation, but instead uses a gradient replacement strategy pioneered by Briggs et al, 2018 Science.doi:10.17504/protocols.io.qmmdu46 fatcat:fmnwcrx67neq3py4prxwce557m