Amplified detection of single base mismatches with the competing-strand assay reveals complex kinetic and thermodynamic behavior of strand displacement at the electrode surface
[2];
- Univ. Rostock (Germany); Univ. at Albany, State Univ. of New York, Albany, NY (United States)
- Univ. at Albany, State Univ. of New York, Albany, NY (United States); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
- Univ. Rostock (Germany)
- Masaryk Univ., Brno (Czech Republic); Institute for Biophysics of the CAS, Brno (Czech Republic)
- Univ. at Albany, State Univ. of New York, Albany, NY (United States); Gensoric GmbH, Rostock (Germany)
Detection of single-base mismatches with respect to a probe strand has been a predominant pursuit in electrochemical biosensor efforts, due to links found between single nucleotide polymorphisms (SNPs) and the predisposition to various diseases. We report an osmium tetroxide bipyridine-based, thermally-controlled, competitive-strand electrochemical assay to allow amplified detection of single-base mismatches. Optimally designed competitive-strand displacement and hybridization temperature allows us to distinguish the single-mismatched-target from the fully complementary sequence with unambiguous, highly reproducible, robust signal differences of over 90%. Furthermore, we find a complex interplay between the position of the redox label, variations in strand displacement kinetics due to mismatches incorporated into the competitive strand, and alterations in the melting temperature of DNA duplexes tethered on the gold surface, when probed by square-wave voltammetry. Furthermore, these insights will apply to any surface-tethered DNA-based electrochemical biosensor, and can help with understanding complex phenomena involved in these types of assays.
- Research Organization:
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Oak Ridge Leadership Computing Facility (OLCF)
- Sponsoring Organization:
- USDOE Office of Science (SC), Advanced Scientific Computing Research (ASCR)
- Grant/Contract Number:
- AC05-00OR22725; AC05-00OR227525; RVO 68081707
- OSTI ID:
- 1464009
- Alternate ID(s):
- OSTI ID: 1702302
- Journal Information:
- Electrochimica Acta, Vol. 285, Issue C; ISSN 0013-4686
- Publisher:
- ElsevierCopyright Statement
- Country of Publication:
- United States
- Language:
- English
Web of Science
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