Quantitation of pyrimidine dimer contents of nonradioactive deoxyribonucleic acid by electrophoresis in alkaline agarose gels.
We have developed a method of quantitating the pyrimidine dimer content of nonradioactive DNAs. DNA samples are treated with the UV-endonuclease from Micrococcus luteus and then separated according to molecular weight by electrophoresis on alkaline agarose gels. From their migration relative to known molecular weight standards, their median molecular weights and thus the number of dimers per DNA molecule in each sample can be calculated. Results of action spectra for dimer formation in T7 bacteriophage measured by this method agree well with action spectra for T7 killing. In addition, the method gives dimer yields in good agreement with those obtained by others using alkaline sucrose gradient sedimentation.
D004269:DNA, Bacterial / Q000302:isolation & purification / Q000528:radiation effects*; D004279:DNA, Viral / Q000302:isolation & purification / Q000528:radiation effects*; D004587:Electrophoresis, Agar Gel / Q000379:methods; D004706:Endodeoxyribonucleases; D004926:Escherichia coli / Q000528:radiation effects*; D007700:Kinetics; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000378:metabolism*; D009699:N-Glycosyl Hydrolases; D011740:Pyrimidine Dimers / Q000032:analysis; D013604:T-Phages / Q000528:radiation effects*; D014466:Ultraviolet Rays
D004269:DNA, Bacterial; D004279:DNA, Viral; D009097:Multienzyme Complexes; D011740:Pyrimidine Dimers; C032219:UV endonuclease; D004706:Endodeoxyribonucleases; D009699:N-Glycosyl Hydrolases