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

Area of Science:

• Molecular Biology
• Genomics
• Bioinformatics

Background:

• High-throughput single-nucleus transcriptomic profiling is crucial for understanding cellular heterogeneity.
• Existing methods for single-cell RNA sequencing can be costly and complex.
• Scalable and cost-effective protocols are needed for broad adoption.

Purpose of the Study:

• To describe an optimized protocol for mouse kidney nuclei isolation and sci-RNA-seq3 library preparation.
• To enable high-throughput, multi-sample transcriptomic profiling.
• To provide a low-cost, scalable method for single-cell RNA sequencing.

Main Methods:

• Optimized protocol for mouse kidney nuclei isolation using a dounce tissue homogenizer for high yield.
• Fixed nuclei processed for sci-RNA-seq3.
• Self-loaded transposome Tn5 used for tagmentation in library generation.

Main Results:

• High yield of nuclei isolated from mouse kidneys.
• Successful generation of scalable single-cell transcriptomic data using sci-RNA-seq3.
• Protocol utilizes common laboratory supplies for cost-effectiveness.

Conclusions:

• The described protocol offers an efficient and cost-effective method for mouse kidney nuclei isolation and sci-RNA-seq3 library preparation.
• This optimized protocol facilitates high-throughput, scalable single-cell transcriptomic data generation.
• Researchers can leverage this protocol with standard laboratory equipment for low-cost transcriptomic analysis.