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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
Chapitres
- 00:05Titre
- 02:53Molecular Beacon Design
- 04:31Nuclear Lysate Preparation
- 08:31Molecular Beacon Assay
- 11:51Representative Analysis of Tumor Cell Lysates
- 13:54Conclusion
Vi beskriver en metode for kvantitativ, real-time måling av DNA glykosylase og AP endonuclease aktiviteter i celle kjernefysiske lysates. Analysen gir tallene for DNA Repair aktivitet mottagelig til kinetisk analyse og er tilpasningsdyktig for kvantifisering av DNA Repair aktivitet i vev og tumor lysates eller med rensede proteiner.
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
