Restriction nuclease digestions driven to completion by Escherichia coli RNA polymerase and T4 gene 32 protein.
Restriction enzyme digestions of large scale DNA preparations often do not go to completion. This is due to product inhibition by the newly generated ends of the digested DNA. The addition of exogenous proteins that bind tightly to the free ends of DNA or to single-stranded DNA will relieve this inhibition. We show that a considerable savings on restriction nucleases can be attained by the addition of RNA polymerase or T4 gene 32 protein in stoichiometric amounts to the newly produced DNA ends.
Dombroski
D F
DF
Morgan
A R
AR
D004262:DNA Restriction Enzymes / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D007700:Kinetics; D010957:Plasmids; D013604:T-Phages / Q000235:genetics*; D014764:Viral Proteins / Q000378:metabolism*
D014764:Viral Proteins; D012321:DNA-Directed RNA Polymerases; D004262:DNA Restriction Enzymes