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Genetics

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JoVE Journal Genetics
A Simple, Rapid, and Quantitative Assay to Measure Repair of DNA-protein Crosslinks on Plasmids Transfected into Mammalian Cells
 

A Simple, Rapid, and Quantitative Assay to Measure Repair of DNA-protein Crosslinks on Plasmids Transfected into Mammalian Cells

Article DOI: 10.3791/57413-v • 11:58 min • March 5th, 2018
March 5th, 2018 •
1, 1
1Department of Pharmacology, University of Minnesota

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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.

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DNA-protein Crosslinks Plasmids Mammalian Cells Quantification Assay DNA Repair Adducts Transfection Trans-Pacific Primer Extension QPCR Essay DNA Damage Response Repair Pathways Abasic Sites Polymerase Blocking Adducts Detection Time Points House Cell Reactivation RNA Polymerase Visual Demonstration Purification Strand-specific Primer Extension
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