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1Department of Pharmacology & Chemical Biology,University of Pittsburgh School of Medicine, 2Hillman Cancer Center,University of Pittsburgh Cancer Institute, 3Department of Experimental Therapy,The Netherlands Cancer Institute, 4Department of Human Genetics,University of Pittsburgh School of Public Health
Capitoli
- 00:05Titolo
- 02:53Molecular Beacon Design
- 04:31Nuclear Lysate Preparation
- 08:31Molecular Beacon Assay
- 11:51Representative Analysis of Tumor Cell Lysates
- 13:54Conclusion
We describe a method for the quantitative, real-time measurement of DNA glycosylase and AP endonuclease activities in cell nuclear lysates. The assay yields rates of DNA Repair activity amenable to kinetic analysis and is adaptable for quantification of DNA Repair activity in tissue and tumor lysates or with purified proteins.
Tags
Quantitative AnalysisReal-time MeasurementBase Excision RepairDNA GlycosylaseAP EndonucleaseMolecular BeaconsSubstrateDeoxyoligonucleotideCarboxyfluorescein (6-FAM)DabcylStem-loop StructureNucleotidesBase PairsFörster Resonance Energy Transfer (FRET)FluorescenceDNA RepairFluorescence ValuesReal-time AssessmentPCR Instruments
