Joining of simian virus 40 DNA molecules at endonuclease R Eco Ri sites by polynucleotide ligase and analysis of the products by agarose gel electrophoresis.
DNA molecules cut with endonuclease R Eco Ri can be joined at Eco Ri cleavage sites by incubation with polynucleotide ligase. In order to define the optimum conditions for this reaction, linear Simian Virus 40 DNA molecules (SV40(Lri)) produced by endonuclease R Eco Ri cleavage of SV40 form i DNA were joined using polynucleotide ligases specified by bacteriophage T4 and Escherichia coli. We have determined that the concentration of the substrate DNA molecules is the most important factor determining the distribution of covalently joined product molecules into a variety of circular and linear monomeric and oligomeric species.
D004262:DNA Restriction Enzymes; D004279:DNA, Viral; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli / Q000201:enzymology; D008970:Molecular Weight; D011118:Polynucleotide Ligases / Q000378:metabolism*; D013539:Simian virus 40
D004279:DNA, Viral; D004262:DNA Restriction Enzymes; D011118:Polynucleotide Ligases