Rapid amperometric verification of PCR amplification of DNA

Thierry De Lumley-Woodyear, Charles N. Campbell, Esti Freeman, Amihay Freeman, George Georgiou, Adam Heller*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

50 Scopus citations


Amplification of an 800-base template was verified in a 10-min test on a 2-μL sample of the PCR product solution. For verification, digoxigeninylated primers and biotinylated d-UTP-16-biotin were added to the amplification solution. The resulting amplified product was digoxigenin-labeled at its 3'- end and was also labeled with multiple biotin functions along its chain. The detecting electrode was coated with an electron-conducting redox hydrogel to which anti-digoxin monoclonal antibody was covalently bound. The amplified DNA was captured by the electrode through conjugation of its 3'-digoxigenin with the antibody. Exposure to a solution of horseradish peroxidase-labeled avidin led to capture of the enzyme and switched the redox hydrogel from a noncatalyst to a catalyst for H2O2 electroreduction. The switching resulted in an H2O2 electroreduction current density of 2.1 ± 0.9 μA cm-2 in 10-4 M H2O2 at Ag/AgCl potential and at 25 °C.

Original languageEnglish
Pages (from-to)535-538
Number of pages4
JournalAnalytical Chemistry
Issue number3
StatePublished - 1 Feb 1999


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