Ultraviolet irradiation of DNA: a way of generating partial digests for rapid restriction site mapping.
UV-irradiation of DNA can inhibit the activity of certain restriction endonucleases because of thymine dimer formation within the enzyme recognition sequence. The number of sites affected depends upon the dose of UV, thus making it easier to control the extent of enzyme digestion than by either limiting the digestion time, or the amount of enzyme. Restriction-site maps of bacteriophage lambda recombinants are readily produced by labelling DNA using a radioactive oligonucleotide that is complementary to either the left or right cohesive end of lambda, irradiating the DNA with UV light, limit digesting with the appropriate enzyme, and calculating the size of the fragments detected after gel electrophoresis and autoradiography.
D010582:Bacteriophage lambda; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D004279:DNA, Viral / Q000528:radiation effects*; D015246:Deoxyribonuclease EcoRI; D004587:Electrophoresis, Agar Gel; D013379:Substrate Specificity; D014466:Ultraviolet Rays
D004279:DNA, Viral; D004262:DNA Restriction Enzymes; D015246:Deoxyribonuclease EcoRI