Rapid mitochondrial DNA isolation method for direct sequencing

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

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

Rapid mitochondrial DNA isolation method for direct sequencing

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

Genetics

Research output: Contribution to journal › Article › peer-review

5 Scopus citations

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 language	English (US)
Pages (from-to)	89-95
Number of pages	7
Journal	Methods in Molecular Biology
Volume	1264
DOIs	https://doi.org/10.1007/978-1-4939-2257-4_9
State	Published - 2015

Keywords

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

ASJC Scopus subject areas

Molecular Biology
Genetics

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

Cite this

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title = "Rapid mitochondrial DNA isolation method for direct sequencing",

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.",

keywords = "Mitochondrial DNA, Next-generation sequencing, Paramagnetic beads, Sequencing libraries, Solid-phase reversible immobilization",

author = "Wilber Quispe and White, {Ryan R.} and Popov, {Vasily N.} and Jan Vijg and Maslov, {Alexander Y.}",

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T1 - Rapid mitochondrial DNA isolation method for direct sequencing

AU - Quispe, Wilber

AU - White, Ryan R.

AU - Popov, Vasily N.

AU - Vijg, Jan

AU - Maslov, Alexander Y.

N1 - Publisher Copyright: © Springer Science+Business Media New York 2015.

PY - 2015

Y1 - 2015

N2 - 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.

AB - 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.

KW - Mitochondrial DNA

KW - Next-generation sequencing

KW - Paramagnetic beads

KW - Sequencing libraries

KW - Solid-phase reversible immobilization

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SP - 89

EP - 95

JO - Methods in Molecular Biology

JF - Methods in Molecular Biology

SN - 1064-3745

ER -

Rapid mitochondrial DNA isolation method for direct sequencing

Abstract

Keywords

ASJC Scopus subject areas

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