SERS-melting: a new method for discriminating mutations in dna sequences
SERS-melting: a new method for discriminating mutations in dna sequences
The reliable discrimination of mutations, single nucleotide polymorphisms (SNPs), and other differences in genomic sequence is an essential part of DNA diagnostics and forensics. It is commonly achieved using fluorescently labeled DNA probes and thermal gradients to distinguish between the matched and mismatched DNA.
Here, we describe a novel method that uses surface enhanced (resonance) Raman spectroscopy (SER(R)S) to follow denaturation of dsDNA attached to a structured gold surface. This denaturation is driven either electrochemically or thermally on SERS active sphere segment void (SSV) gold substrates. Using this method, we can distinguish between wild type, a single point mutation (1653C/T), and a triple deletion (?F 508) in the CFTR gene at the 0.02 attomole level, and the method can be used to differentiate the unpurified PCR products of the wild type and ?F 508 mutation.
Our method has the potential to provide small, rapid, sensitive, reproducible platforms for detecting genetic variations and sequencing genes.
15589-15601
Mahajan, Sumeet
b131f40a-479e-4432-b662-19d60d4069e9
Richardson, James
51db9f73-a136-48af-8722-21d46906cf40
Brown, Tom
a64aae36-bb30-42df-88a2-11be394e8c89
Bartlett, Philip N.
d99446db-a59d-4f89-96eb-f64b5d8bb075
19 November 2008
Mahajan, Sumeet
b131f40a-479e-4432-b662-19d60d4069e9
Richardson, James
51db9f73-a136-48af-8722-21d46906cf40
Brown, Tom
a64aae36-bb30-42df-88a2-11be394e8c89
Bartlett, Philip N.
d99446db-a59d-4f89-96eb-f64b5d8bb075
Mahajan, Sumeet, Richardson, James, Brown, Tom and Bartlett, Philip N.
(2008)
SERS-melting: a new method for discriminating mutations in dna sequences.
Journal of the American Chemical Society, 130 (46), .
(doi:10.1021/ja805517q).
Abstract
The reliable discrimination of mutations, single nucleotide polymorphisms (SNPs), and other differences in genomic sequence is an essential part of DNA diagnostics and forensics. It is commonly achieved using fluorescently labeled DNA probes and thermal gradients to distinguish between the matched and mismatched DNA.
Here, we describe a novel method that uses surface enhanced (resonance) Raman spectroscopy (SER(R)S) to follow denaturation of dsDNA attached to a structured gold surface. This denaturation is driven either electrochemically or thermally on SERS active sphere segment void (SSV) gold substrates. Using this method, we can distinguish between wild type, a single point mutation (1653C/T), and a triple deletion (?F 508) in the CFTR gene at the 0.02 attomole level, and the method can be used to differentiate the unpurified PCR products of the wild type and ?F 508 mutation.
Our method has the potential to provide small, rapid, sensitive, reproducible platforms for detecting genetic variations and sequencing genes.
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e-pub ahead of print date: 28 October 2008
Published date: 19 November 2008
Organisations:
Chemistry, Institute for Life Sciences
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Local EPrints ID: 145029
URI: http://eprints.soton.ac.uk/id/eprint/145029
ISSN: 0002-7863
PURE UUID: c7d12290-745d-4f39-9d3b-a9e0507ecca9
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Date deposited: 15 Apr 2010 14:42
Last modified: 14 Mar 2024 02:52
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Author:
James Richardson
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