Issue 1, 2011

SPR-based single nucleotide mismatch biosensor

Abstract

The detection and characterization of the hybridization event of 21-base, unlabeled DNA oligonucleotides with a monolayer of complementary DNA immobilized on a gold surface, by electrochemical impedance spectroscopy and surface plasmon resonance (SPR) is presented. A thiol modification on the probe DNA strand allowed for its attachment to the surface viaself-assembly. For the hybridization of full match target DNA a detection limit of 20 pM was determined. RNA hybridization was also detectable with the same sensor, with a similar detection limit. The SPR signal generated upon hybridization of the full match was always distinguishable from the single mismatch target DNA oligonucleotides when the mismatch was in the middle or at the proximal end of the target DNA sequence. However, the response of the sensor was identical for the hybridization of the full match and the distal end mismatch. The SPR sensor described is reusable over at least 20 hybridization/regeneration cycles and is insensitive to flow rate (20–800 µL min−1) or temperature (20–60 °C). Based on the SPR response, the surface density of the probe was estimated to be at least 4.3 × 1012 molecules per cm2.

Graphical abstract: SPR-based single nucleotide mismatch biosensor

Article information

Article type
Paper
Submitted
10 Aug 2010
Accepted
24 Sep 2010
First published
23 Nov 2010

Anal. Methods, 2011,3, 122-132

SPR-based single nucleotide mismatch biosensor

E. Milkani, A. M. Khaing, S. Morais, C. R. Lambert and W. G. McGimpsey, Anal. Methods, 2011, 3, 122 DOI: 10.1039/C0AY00492H

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