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Rapid Mitochondrial DNA Isolation Method for Direct Sequencing

  • Wilber Quispe-Tintaya
  • Ryan R. White
  • Vasily N. Popov
  • Jan Vijg
  • Alexander Y. MaslovEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1264)

Abstract

Standard methods for mitochondrial DNA (mtDNA) extraction do not provide the level of enrichment for mtDNA sufficient for direct sequencing and must be followed by long-range-PCR amplification, which can bias the sequence results. Here, we describe a reliable method for the preparation of mtDNA-enriched samples from eukaryotic cells ready for direct sequencing. This protocol utilizes a conventional miniprep kit, in conjunction with a paramagnetic bead-based purification step.

Key words

Next-generation sequencing Mitochondrial DNA Paramagnetic beads Solid-phase reversible immobilization Sequencing libraries 

Notes

Acknowledgment

This research was supported by NIH grant P01 AG017242 (JV) and by the Ministry of Education and Science of Russian Federation grant 14.B37.21.1966 (VNP and AYM).

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Wilber Quispe-Tintaya
    • 1
  • Ryan R. White
    • 1
  • Vasily N. Popov
    • 2
  • Jan Vijg
    • 1
  • Alexander Y. Maslov
    • 1
    Email author
  1. 1.Department of GeneticsAlbert Einstein College of MedicineNew YorkUSA
  2. 2.Department of Genetics, Cytology, and BioengineeringVoronezh State UniversityVoronezhRussia

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