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Measuring Single-Cell Mitochondrial DNA Copy Number and Heteroplasmy Using Digital Droplet Polymerase Chain Reaction
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Measuring Single-Cell Mitochondrial DNA Copy Number and Heteroplasmy Using Digital Droplet Polymerase Chain Reaction

Measuring Single-Cell Mitochondrial DNA Copy Number and Heteroplasmy Using Digital Droplet Polymerase Chain Reaction

DOI:
10.3791/63870-v

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09:15 min

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July 12, 2022

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1Department of Clinical Neurosciences, School of Clinical Medicine,University of Cambridge, Cambridge Biomedical Campus, 2Medical Research Council Mitochondrial Biology Unit,University of Cambridge, Cambridge Biomedical Campus

Chapters

  • 00:04Introduction
  • 00:52Preparation of Samples
  • 01:51Generation of Droplets
  • 04:16PCR and Droplet Reading
  • 06:07Analysis of Results
  • 07:13Results: Measuring Mitochondrial DNA Copy Number and Heteroplasmy in Single Cells
  • 08:28Conclusion

Summary

Automatic Translation

Automatic Translation

Here we present a protocol for measuring absolute mitochondrial (mt)DNA copy number and mtDNA deletion heteroplasmy levels in single cells.

Tags

Keywords: Mitochondrial DNASingle-cellDigital Droplet PCRHeteroplasmyCopy NumberCell LysisSingle-cell SortingMaster MixDroplet GenerationNon-template Control

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