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Protocol for immunodot blot detection of UVB photoproducts in extracellular DNA
M Alexandra Carpenter1, Sri Yerrapragada1, Aleena Alex1
1Department of Pharmacology & Toxicology, Wright State University Boonshoft School of Medicine, Dayton, OH 45435, USA.
Abstract:
UV radiation induces the formation of adducts in genomic DNA within cells that are later found to be present in cell-free fractions associated with extracellular vesicles (EVs) outside of cells. Here, we present a protocol for isolating UV photoproducts in extracellular DNA released from UVB-irradiated cells via differential centrifugation. We then detail steps for monitoring the DNA adducts using DNA immunoblotting. This protocol can be applied for detection of DNA adducts in EVs from cell culture and skin explant models. For complete details on the use and execution of this protocol, please refer to Carpenter et al.1.