Postdoctoral fellow, Oak Ridge Associated Universities, Oak Ridge, TN, USA.
Matrix-assisted laser desorption/ionization detection of polymerase chain reaction products by utilizing the 5′-3′ exonuclease activity of Thermus aquaticus DNA polymerase†
Article first published online: 11 FEB 2003
Copyright © 2003 John Wiley & Sons, Ltd.
Rapid Communications in Mass Spectrometry
Volume 17, Issue 6, pages 532–537, 30 March 2003
How to Cite
Isola, N. R., Liu, Z., Allman, S. L., Taranenko, N. I., Kong, Y. and Chen, C. H. (2003), Matrix-assisted laser desorption/ionization detection of polymerase chain reaction products by utilizing the 5′-3′ exonuclease activity of Thermus aquaticus DNA polymerase. Rapid Commun. Mass Spectrom., 17: 532–537. doi: 10.1002/rcm.939
The submitted manuscript has been authored by a contractor of the U.S. Government under contract No. DE-AC05-00OR22725. Accordingly, the U.S. Government retains a nonexclusive, royalty-free license to publish or reproduce the published form of this contribution, or allow others to do so, for U.S. Government purposes.
- Issue published online: 11 FEB 2003
- Article first published online: 11 FEB 2003
- Manuscript Accepted: 3 JAN 2003
- Manuscript Revised: 14 DEC 2002
- Manuscript Received: 9 OCT 2002
The 5′-3′ exonuclease activity of DNA polymerase was utilized in the polymerase chain reaction system to generate a specific signal concomitant with amplification. These signals were detected by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). This method obviates the need to perform extensive DNA purification of reaction products that is often necessary for detecting larger DNA molecules by mass spectrometry. Oligonucleotides complementary to the internal region of the amplicon are degraded by the 5′-3′ exonuclease activity and the degradation products are analyzed by MALDI mass spectrometry. We refer to this assay as the Exo-taq assay or probe degradation assay. This method should be amenable to automation. Copyright © 2003 John Wiley & Sons, Ltd.