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


Li Haikuo and Humphreys Benjamin D.

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.

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Tagmentase

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Published
December, 2022

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Products used in this publication

  • Tubes
    C01070010-10
    Tagmentase (Tn5 transposase) - unloaded

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