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 | |
| State | 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
Rapid mitochondrial DNA isolation method for direct sequencing. / Quispe-Tintaya, Wilber; White, Ryan R.; Popov, Vasily N.; Vijg, Jan; Maslov, Alexander.
Probing Mitochondrial Function. Vol. 1 Springer New York, 2015. p. 89-95.Research output: Chapter in Book/Report/Conference proceeding › Chapter
}
TY - CHAP
T1 - Rapid mitochondrial DNA isolation method for direct sequencing
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.
KW - Mitochondrial DNA
KW - Next-generation sequencing
KW - Paramagnetic beads
KW - Sequencing libraries
KW - Solid-phase reversible immobilization
UR - http://www.scopus.com/inward/record.url?scp=84958590781&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84958590781&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-2257-4_9
DO - 10.1007/978-1-4939-2257-4_9
M3 - Chapter
C2 - 25631006
AN - SCOPUS:84958590781
SN - 9781493922574
SN - 9781493922567
VL - 1
SP - 89
EP - 95
BT - Probing Mitochondrial Function
PB - Springer New York
ER -