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Temporal temperature gradient electrophoresis for detection of single nucleotide polymorphisms

Temporal temperature gradient electrophoresis for detection of single nucleotide polymorphisms
Temporal temperature gradient electrophoresis for detection of single nucleotide polymorphisms
The presence of single nucleotide polymorphisms (SNPs) in nuclear DNA and mitochondrial DNA (mtDNA) can be detected using a range of electrophoretic techniques, of which temporal temperature gradient electrophoresis (TTGE) is often the most user-friendly and reproducible. The technique operates on the same principle as denaturing gradient gel electrophoresis, but does not require a chemical gradient in the gel. Instead, TTGE relies on a steady and gradual increase in temperature during electrophoresis to denature and separate DNA sequences that differ by as little as one base pair. TTGE can be easily accomplished using DNA of high quality and it is a rapid-throughput method for SNP screening once conditions have been optimized. Detection of SNPs is, for example, important for the diagnosis of mitochondrial disorders such as heteroplasmy, the presence of more than one type of mitochondria within a cell or tissue. Here we describe the basic steps for TTGE and illustrate its utility for the detection of heteroplasmy in mtDNA control region sequences.

1064-3745
153-165
Jones, Bethan M.
2a40cabe-e5ee-4454-8aab-585c3af9fa70
Knapp, Leslie A.
1b4bc671-2fc7-47f7-bccb-231fbd5cfd20
Jones, Bethan M.
2a40cabe-e5ee-4454-8aab-585c3af9fa70
Knapp, Leslie A.
1b4bc671-2fc7-47f7-bccb-231fbd5cfd20

Jones, Bethan M. and Knapp, Leslie A. (2009) Temporal temperature gradient electrophoresis for detection of single nucleotide polymorphisms. Methods in Molecular Biology, 578, 153-165.

Record type: Article

Abstract

The presence of single nucleotide polymorphisms (SNPs) in nuclear DNA and mitochondrial DNA (mtDNA) can be detected using a range of electrophoretic techniques, of which temporal temperature gradient electrophoresis (TTGE) is often the most user-friendly and reproducible. The technique operates on the same principle as denaturing gradient gel electrophoresis, but does not require a chemical gradient in the gel. Instead, TTGE relies on a steady and gradual increase in temperature during electrophoresis to denature and separate DNA sequences that differ by as little as one base pair. TTGE can be easily accomplished using DNA of high quality and it is a rapid-throughput method for SNP screening once conditions have been optimized. Detection of SNPs is, for example, important for the diagnosis of mitochondrial disorders such as heteroplasmy, the presence of more than one type of mitochondria within a cell or tissue. Here we describe the basic steps for TTGE and illustrate its utility for the detection of heteroplasmy in mtDNA control region sequences.

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Published date: 2009

Identifiers

Local EPrints ID: 69257
URI: http://eprints.soton.ac.uk/id/eprint/69257
ISSN: 1064-3745
PURE UUID: 3823a03e-de98-41cf-8a0f-5a573ea8a07d

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Date deposited: 26 Oct 2009
Last modified: 27 Apr 2022 08:31

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Contributors

Author: Bethan M. Jones
Author: Leslie A. Knapp

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