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Ethidium-bromide-inhibited restriction endonucleases cleave one strand of circular DNA
Authors:M Osterlund  H Luthman  S V Nilsson  G Magnusson
Affiliation:The Medical Nobel Institute, Department of Biochemistry I, Karolinska Institute, S-104 01 StockholmSweden
Abstract:We analyzed the effect of ethidium bromide (EtBr) on the cleavage of closed circular pBR322 DNA molecules by six restriction enzymes which make staggered or flush cuts (EcoRI, HindIII, BglI, PstI, HincII, PvuII). EtBr concentrations and reaction temperatures were determined at which DNA molecules with single-strand breaks were the major reaction product of digestion by all the enzymes. However, the amounts of intermediates which could be isolated differed for various enzymes. The results extend previous studies, showing that sequential cleavage of the DNA strands probably is a general property of restriction endonucleases.
Keywords:Intercalation  plasmid pBR322  increased dye binding of cleavage intermediates  stabilization of cleavage intermediates  DNA I  covalently closed circular of DNA  DNA II  open circular of DNA  DNA III  linear circular forms of DNA  EtBr  ethidium bromide  UV  ultraviolet
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