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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
Beskriver vi en fremgangsmåde til kvantitativ real-time måling af DNA-glycosylase og AP-endonuclease aktiviteter cellekerner lysater. Assayet giver hastigheder af DNA-reparation aktivitet kan underkastes kinetisk analyse og kan tilpasses til kvantificering af DNA-reparation aktivitet i væv og tumorer lysater eller oprensede 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
