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

4 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
Publication status	Published - Jan 28 2015

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Keywords

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

ASJC Scopus subject areas

Biochemistry, Genetics and Molecular Biology(all)

Cite this

Quispe-Tintaya, W., White, R. R., Popov, V. N., Vijg, J., & Maslov, A. (2015). Rapid mitochondrial DNA isolation method for direct sequencing. In Probing Mitochondrial Function (Vol. 1, pp. 89-95). Springer New York. https://doi.org/10.1007/978-1-4939-2257-4_9

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AU - Maslov, Alexander

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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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Access to Document

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