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A Simple, Rapid, and Quantitative Assay to Measure Repair of DNA-protein Crosslinks on Plasmids Transfected into Mammalian Cells
Capítulos
- 00:04Título
- 01:18Generation of DNA-protein Crosslink Containing Plasmid
- 06:21Transfection into Mammalian Cells
- 08:39SSPE-qPCR
- 10:02Results: Use of Strand-specific Primer Extension Based q-PCR to Quantify the DNA-protein Crosslinks
- 11:08Conclusion
The goal of this protocol is to quantify the repair of defined DNA-protein crosslinks on plasmid DNA. Lesioned plasmids are transfected into recipient mammalian cell lines and low-molecular weight harvested at multiple time points post-transfection. DNA repair kinetics are quantified using strand-specific primer extension followed by qPCR.
