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A slot blot method for detection of ultraviolet photoproducts in DNA
1Department of Biochemistry and Biophysics, Washington State University, Pullman 99164-4660, USA.
Analytical Biochemistry
|August 10, 1995
Abstract:
A simple new method was established to detect the major uv photoproducts in DNA. A slot blot method, involving the use of T4 DNA polymerase-associated (3'--> 5') exonuclease digestion of uv-irradiated DNA, was used to detect (primarily) cyclobutane pyrimidine dimers and pyrimidine-pyrimidone (6-4) dimers. Hydrolysis of DNA by this enzyme is quantitatively blocked by both of these photoproducts. To detect (primarily) (6-4) dimers, Escherichia coli DNA photolyase was utilized to specifically reverse cyclobutane pyrimidine dimers.