Nonequilibrium Hybridization Enables Discrimination Of A Point Mutation Within 5-40 °C

Abstract

Detection of point mutations and single nucleotide polymorphisms in DNA and RNA has a growing importance in biology, biotechnology, and medicine. For the application at hand, hybridization assays are often used. Traditionally, they differentiate point mutations only at elevated temperatures (>40 °C) and in narrow intervals (ΔT = 1-10 °C). The current study demonstrates that a specially designed multistranded DNA probe can differentiate point mutations in the range of 5-40 °C. This unprecedentedly broad ambient-temperature range is enabled by a controlled combination of (i) nonequilibrium hybridization conditions and (ii) a mismatch-induced increase of equilibration time in respect to that of a fully matched complex, which we dub "kinetic inversion".

Publication Date

10-19-2016

Publication Title

Journal of the American Chemical Society

Volume

138

Issue

41

Number of Pages

13465-13468

Document Type

Article

Personal Identifier

scopus

DOI Link

https://doi.org/10.1021/jacs.6b05628

Socpus ID

84992213204 (Scopus)

Source API URL

https://api.elsevier.com/content/abstract/scopus_id/84992213204

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