Rapid mitochondrial DNA isolation method for direct sequencing

Wilber Quispe-tintaya, Ryan R. White, Vasily N. Popov, Jan Vijg, Alexander Maslov

Research output: Contribution to journalArticle

Abstract

Standard methods for mitochondrial DNA (mtDNA) extraction do not provide the level of enrichment for mtDNA suffi cient for direct sequencing and must be followed by long-range-PCR amplifi cation, 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 purifi cation step.

Original languageEnglish (US)
Pages (from-to)89-95
Number of pages7
JournalMethods in molecular biology (Clifton, N.J.)
Volume1264
DOIs
StatePublished - 2015
Externally publishedYes

Fingerprint

Mitochondrial DNA
Cations
Eukaryotic Cells
Polymerase Chain Reaction

Keywords

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

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Rapid mitochondrial DNA isolation method for direct sequencing. / Quispe-tintaya, Wilber; White, Ryan R.; Popov, Vasily N.; Vijg, Jan; Maslov, Alexander.

In: Methods in molecular biology (Clifton, N.J.), Vol. 1264, 2015, p. 89-95.

Research output: Contribution to journalArticle

Quispe-tintaya, Wilber ; White, Ryan R. ; Popov, Vasily N. ; Vijg, Jan ; Maslov, Alexander. / Rapid mitochondrial DNA isolation method for direct sequencing. In: Methods in molecular biology (Clifton, N.J.). 2015 ; Vol. 1264. pp. 89-95.
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