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Mouse kidney nuclear isolation and library preparation for single-cell combinatorial indexing RNA sequencing.

ABSTRACT: Single-cell combinatorial indexing RNA sequencing (sci-RNA-seq3) enables high-throughput single-nucleus transcriptomic profiling of multiple samples in one experiment. Here, we describe an optimized protocol of mouse kidney nuclei isolation and sci-RNA-seq3 library preparation. The use of a dounce tissue homogenizer enables nuclei extraction with high yield. Fixed nuclei are processed for sci-RNA-seq3, and self-loaded transposome Tn5 is used for tagmentation in library generation. The step-by-step protocol allows researchers to generate scalable single-cell transcriptomic data with common laboratory supplies at low cost. For complete details on the use and execution of this protocol, please refer to Li et al. (2022).¹.

SUBMITTER: Li H

PROVIDER: S-EPMC9732400 | biostudies-literature | 2022 Dec

REPOSITORIES: biostudies-literature

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Mouse kidney nuclear isolation and library preparation for single-cell combinatorial indexing RNA sequencing.

Li Haikuo H Humphreys Benjamin D BD

STAR protocols 20221205 4

Single-cell combinatorial indexing RNA sequencing (sci-RNA-seq3) enables high-throughput single-nucleus transcriptomic profiling of multiple samples in one experiment. Here, we describe an optimized protocol of mouse kidney nuclei isolation and sci-RNA-seq3 library preparation. The use of a dounce tissue homogenizer enables nuclei extraction with high yield. Fixed nuclei are processed for sci-RNA-seq3, and self-loaded transposome Tn5 is used for tagmentation in library generation. The step-by-st ...[more]

PMID: 36595916

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