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HERO ID
2879659
Reference Type
Journal Article
Title
DNA extraction procedures meaningfully influence qPCR-based mtDNA copy number determination
Author(s)
Guo, W; Jiang, L; Bhasin, S; Khan, SM; Swerdlow, RH
Year
2009
Is Peer Reviewed?
1
Journal
Mitochondrion
ISSN:
1567-7249
Volume
9
Issue
4
Page Numbers
261-265
Language
English
PMID
19324101
DOI
10.1016/j.mito.2009.03.003
Web of Science Id
WOS:000267118000006
Abstract
Quantitative real time PCR (qPCR) is commonly used to determine cell mitochondrial DNA (mtDNA) copy number. This technique involves obtaining the ratio of an unknown variable (number of copies of an mtDNA gene) to a known parameter (number of copies of a nuclear DNA gene) within a genomic DNA sample. We considered the possibility that mtDNA:nuclear DNA (nDNA) ratio determinations could vary depending on the method of genomic DNA extraction used, and that these differences could substantively impact mtDNA copy number determination via qPCR. To test this we measured mtDNA:nDNA ratios in genomic DNA samples prepared using organic solvent (phenol-chloroform-isoamyl alcohol) extraction and two different silica-based column methods, and found mtDNA:nDNA ratio estimates were not uniform. We further evaluated whether different genomic DNA preparation methods could influence outcomes of experiments that use mtDNA:nDNA ratios as endpoints, and found the method of genomic DNA extraction can indeed alter experimental outcomes. We conclude genomic DNA sample preparation can meaningfully influence mtDNA copy number determination by qPCR.
Keywords
Mitochondrial DNA; Phenol extraction; Real-times PCR; DNA isolation
Tags
•
Chloroform 2018 Update
TITLE AND ABSTRACT REVIEW
Excluded Studies
•
Chloroform Combined (current)
Chloroform (original)
References: 2000-2018
WoS
Chloroform (2018 update)
TITLE AND ABSTRACT REVIEW
Excluded Studies
Chloroform (current)
Literature Search: Jan 2009 - March 2017
Web of Science
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