Single-base mismatch detection based on charge transduction through DNA

Shana O. Kelley, Elizabeth M. Boon, Jacqueline K. Barton*, Nicole M. Jackson, Michael G. Hill

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

515 Scopus citations

Abstract

High-throughput DNA sensors capable of detecting single-base mismatches are required for the routine screening of genetic mutations and disease. A new strategy for the electrochemical detection of single-base mismatches in DNA has been developed based upon charge transport through DNA films. Double-helical DNA films on gold surfaces have been prepared and used to detect DNA mismatches electrochemically. The signals obtained from redox-active intercalators bound to DNA-modified gold surfaces display a marked sensitivity to the presence of base mismatches within the immobilized duplexes. Differential mismatch detection was accomplished irrespective of DNA sequence composition and mismatch identity. Single-base changes in sequences hybridized at the electrode surface are also detected accurately. Coupling the redox reactions of intercalated species to electrocatalytic processes in solution considerably increases the sensitivity of this assay. Reporting on the electronic structure of DNA, as opposed to the hybridization energetics of single-stranded oligonucleotides, electrochemical sensors based on charge transport may offer fundamental advantages in both scope and sensitivity.

Original languageEnglish (US)
Pages (from-to)4830-4837
Number of pages8
JournalNucleic acids research
Volume27
Issue number24
DOIs
StatePublished - Dec 15 1999
Externally publishedYes

ASJC Scopus subject areas

  • Genetics

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