A Protocol for Single Nucleus RNAseq from Frozen Skeletal Muscle

Author:

Soule Tyler GB,Pontifex Carly S,Rosin Nicole,Joel Matthew M,Lee Sukyoung,Nguyen Minh Dang,Chhibber Sameer,Pfeffer GeraldORCID

Abstract

AbstractSingle cell technologies are a method of choice to obtain vast amounts of cell-specific transcriptional information under physiological and diseased states. Myogenic cells are resistant to single nucleus RNA sequencing (snRNAseq) due to their large, multinucleated nature. Here, we report a novel, reliable, and cost-effective method to analyze frozen human skeletal muscle by snRNAseq. This method yields all expected cell types for human skeletal muscle and works on tissue frozen for long periods of time and with significant pathological changes. Our method is ideal for studying banked samples with the intention of studying human muscle disease.

Publisher

Cold Spring Harbor Laboratory

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