Unit

UNIT 19.8 Next Generation Sequencing to Characterize Mitochondrial Genomic DNA Heteroplasmy

  1. Taosheng Huang

Published Online: 1 OCT 2011

DOI: 10.1002/0471142905.hg1908s71

Current Protocols in Human Genetics

Current Protocols in Human Genetics

How to Cite

Huang, T. 2011. Next Generation Sequencing to Characterize Mitochondrial Genomic DNA Heteroplasmy. Current Protocols in Human Genetics. 71:19.8:19.8.1–19.8.12.

Author Information

  1. Division of Human Genetics/Department of Pediatrics, Center for Molecular and Mitochondrial Medicine and Genetics, University of California, Irvine, Irvine, California

Publication History

  1. Published Online: 1 OCT 2011
  2. Published Print: OCT 2011

Abstract

This protocol describes the methodology to characterize mitochondrial DNA (mtDNA) heteroplasmy by parallel sequencing. Mitochondria play an important role in essential cellular functions. Each eukaryotic cell contains hundreds of mitochondria with hundreds of mitochondria genomes. Mutant and wild-type mtDNA may co-exist as heteroplasmy, and cause human disease. The purpose of this protocol is to simultaneously determine mtDNA sequence and quantify the heteroplasmic level. This protocol includes a two-fragment mitochondrial genome DNA PCR amplification. The PCR product is then mixed at an equimolar ratio. The samples are then barcoded and sequenced with high-throughput, next-generation sequencing technology. This technology is highly sensitive, specific, and accurate in determining mtDNA mutations and the level of heteroplasmy. Curr. Protoc. Hum. Genet. 71:19.8.1-19.8.12 © 2011 by John Wiley & Sons, Inc.

Keywords:

  • mitochondria;
  • next-generation sequencing;
  • heteroplasmy