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Cryosectioning of Contiguous Regions of a Single Mouse Skeletal Muscle for Gene Expression and Histological Analyses
Published on: December 12, 2016
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A protocol for single nucleus RNA-seq from frozen skeletal muscle
Tyler Gb Soule1, Carly S Pontifex1, Nicole Rosin1,2
1Hotchkiss Brain Institute, University of Calgary, Calgary, Canada.
Life Science Alliance
|March 13, 2023
Summary
We developed a cost-effective single-nucleus RNA sequencing method for analyzing frozen human skeletal muscle. This approach successfully captures diverse cell types, even in long-term banked or pathologically altered samples, aiding muscle disease research.
Area of Science:
- Molecular Biology
- Genomics
- Biotechnology
Background:
- Single-cell technologies provide valuable cell-specific transcriptional data.
- Myogenic cells present challenges for traditional single-cell RNA sequencing due to their large, multinucleated structure.
Purpose of the Study:
- To develop a novel, reliable, and cost-effective method for analyzing frozen human skeletal muscle using single-nucleus RNA sequencing.
- To enable the study of banked skeletal muscle samples, including those with pathological changes.
Main Methods:
- Single-nucleus RNA sequencing was optimized for frozen human skeletal muscle tissue.
- The method was validated on samples with varying storage durations and pathological conditions.
Main Results:
- The developed method successfully identified all expected cell types in human skeletal muscle.
- The technique proved effective on tissue frozen for extended periods and exhibiting significant pathological alterations.
Conclusions:
- This single-nucleus RNA sequencing method offers a robust solution for analyzing banked human skeletal muscle.
- It is particularly suitable for investigating human muscle diseases using preserved tissue samples.
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