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Specifically horizontally tethered DNA probes on Au surfaces allow labelled and label-free DNA detection using SERS and electrochemically driven melting

Specifically horizontally tethered DNA probes on Au surfaces allow labelled and label-free DNA detection using SERS and electrochemically driven melting
Specifically horizontally tethered DNA probes on Au surfaces allow labelled and label-free DNA detection using SERS and electrochemically driven melting
Controlled covalent attachment of dsDNA horizontally orientated on a gold surface is achieved through the use of a single surface-linker located approximately half way along the attached DNA probe strand. We show that horizontally oriented dsDNA on a gold surface can undergo melting and re-hybridization to target strand in solution and thus can be used for the detection of specific target DNA sequences using surface-enhanced Raman spectroscopy (SERS). We show that a range of lengths of target DNA sequences from ?30-bases to 78-bases can be specifically hybridized to the short immobilized DNA probe sequence and adopt a horizontal orientation on the gold surface. Following thermal or electrochemically driven melting of the immobilized dsDNA, the target DNA strand diffuses away while the probe strand remains attached to the surface allowing the functionalized surfaces to be reused. The melting of the horizontally orientated immobilized dsDNA can be monitored using SERS either by employing a dye label covalently attached on the DNA target strand or by employing a binding agent selective for dsDNA. This approach of covalently immobilizing the DNA probe strand through a linker located at approximately the middle of the strand has great potential to improve the sensitivity and specificity of molecular assays that employ DNA arrays on solid surfaces.
1478-6524
386-393
Papadopoulou, E.
235d2d8f-e2af-4fc0-a402-5fc36184f621
Gale, N.
1a74d27d-b323-4a79-8240-5718c03717e9
Thompson, J.F.
3b2c67fd-c351-4226-9222-2c6ee83b72a9
Fleming, T.A.
31d13a5f-e16b-4f5d-90eb-451ed28e32fa
Brown, T.
65b220ab-5839-4e03-b923-97694339baaf
Bartlett, P.N.
d99446db-a59d-4f89-96eb-f64b5d8bb075
Papadopoulou, E.
235d2d8f-e2af-4fc0-a402-5fc36184f621
Gale, N.
1a74d27d-b323-4a79-8240-5718c03717e9
Thompson, J.F.
3b2c67fd-c351-4226-9222-2c6ee83b72a9
Fleming, T.A.
31d13a5f-e16b-4f5d-90eb-451ed28e32fa
Brown, T.
65b220ab-5839-4e03-b923-97694339baaf
Bartlett, P.N.
d99446db-a59d-4f89-96eb-f64b5d8bb075

Papadopoulou, E., Gale, N., Thompson, J.F., Fleming, T.A., Brown, T. and Bartlett, P.N. (2016) Specifically horizontally tethered DNA probes on Au surfaces allow labelled and label-free DNA detection using SERS and electrochemically driven melting. Chemical Science, 7 (1), 386-393. (doi:10.1039/C5SC03185K).

Record type: Article

Abstract

Controlled covalent attachment of dsDNA horizontally orientated on a gold surface is achieved through the use of a single surface-linker located approximately half way along the attached DNA probe strand. We show that horizontally oriented dsDNA on a gold surface can undergo melting and re-hybridization to target strand in solution and thus can be used for the detection of specific target DNA sequences using surface-enhanced Raman spectroscopy (SERS). We show that a range of lengths of target DNA sequences from ?30-bases to 78-bases can be specifically hybridized to the short immobilized DNA probe sequence and adopt a horizontal orientation on the gold surface. Following thermal or electrochemically driven melting of the immobilized dsDNA, the target DNA strand diffuses away while the probe strand remains attached to the surface allowing the functionalized surfaces to be reused. The melting of the horizontally orientated immobilized dsDNA can be monitored using SERS either by employing a dye label covalently attached on the DNA target strand or by employing a binding agent selective for dsDNA. This approach of covalently immobilizing the DNA probe strand through a linker located at approximately the middle of the strand has great potential to improve the sensitivity and specificity of molecular assays that employ DNA arrays on solid surfaces.

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Accepted/In Press date: 8 October 2015
e-pub ahead of print date: 8 October 2015
Published date: January 2016
Organisations: Electrochemistry

Identifiers

Local EPrints ID: 385446
URI: http://eprints.soton.ac.uk/id/eprint/385446
ISSN: 1478-6524
PURE UUID: 9833686d-a731-4aa6-9d7e-c80cdd7852ae
ORCID for P.N. Bartlett: ORCID iD orcid.org/0000-0002-7300-6900

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Date deposited: 20 Jan 2016 08:59
Last modified: 15 Mar 2024 02:44

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Contributors

Author: E. Papadopoulou
Author: N. Gale
Author: J.F. Thompson
Author: T.A. Fleming
Author: T. Brown
Author: P.N. Bartlett ORCID iD

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