Matrix-assisted laser desorption/ionization detection of polymerase chain reaction products by utilizing the 5?-3? exonuclease activity ofThermus aquaticus DNA polymerase
| Autor: | N. I. Taranenko, Y. Kong, C. H. Chen, Steve L. Allman, Z. Liu, Narayana R. Isola |
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| Rok vydání: | 2003 |
| Předmět: |
Exonucleases
Exonuclease Chromatography Thermus aquaticus biology Chemistry Oligonucleotide DNA polymerase Organic Chemistry DNA biology.organism_classification Mass spectrometry Polymerase Chain Reaction Analytical Chemistry Matrix-assisted laser desorption/ionization Polymerase chain reaction optimization Biochemistry Multienzyme Complexes Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization biology.protein Humans Taq Polymerase Spectroscopy Polymerase |
| Zdroj: | Rapid Communications in Mass Spectrometry. 17:532-537 |
| ISSN: | 1097-0231 0951-4198 |
| DOI: | 10.1002/rcm.939 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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