Rapid mitochondrial DNA isolation method for direct sequencing

Wilber Quispe-Tintaya; Ryan R. White; Vasily N. Popov; Jan Vijg; Alexander Maslov

doi:10.1007/978-1-4939-2257-4_9

Rapid mitochondrial DNA isolation method for direct sequencing

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

Genetics

Research output: Chapter in Book/Report/Conference proceeding › Chapter

5 Scopus citations

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.

Original language	English (US)
Title of host publication	Probing Mitochondrial Function
Publisher	Springer New York
Pages	89-95
Number of pages	7
Volume	1
ISBN (Print)	9781493922574, 9781493922567
DOIs	https://doi.org/10.1007/978-1-4939-2257-4_9
State	Published - Jan 28 2015

Keywords

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

ASJC Scopus subject areas

Biochemistry, Genetics and Molecular Biology(all)

Access to Document

10.1007/978-1-4939-2257-4_9

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AU - Quispe-Tintaya, Wilber

AU - White, Ryan R.

AU - Popov, Vasily N.

AU - Vijg, Jan

AU - Maslov, Alexander

PY - 2015/1/28

Y1 - 2015/1/28

N2 - 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.

AB - 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.

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KW - Next-generation sequencing

KW - Paramagnetic beads

KW - Sequencing libraries

KW - Solid-phase reversible immobilization

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