Click publication title for the full text. Please click the arrow on the right to expand the citation list. Incubation of 10 units of Klenow with supercoiled plasmid DNA produced no nicked molecules after 20 hours at 37 ☌ as determined by agarose gel electrophoresis analysis. Klenow Fragment of DNA Polymerase I from Escherichia coZi (Received for publication, March 30, 1990) Andrea H. The enzyme is greater than 98 % pure as indicated by SDS-polyacrylamide gel electrophoresis and contains no detected endonuclease activity. Dissolve 0.1 - 4 μg of digested DNA in 1x Reaction Buffer supplemented with 40 μM each dNTPĥ00 mM Tris-HCl pH 7.6 at 25 ☌, 50 mM MgCl 2 and 10 mM DTT.Įxcessive amounts of enzyme or longer reaction times may result in recessed ends due to the 3'→5' exonuclease activity of the enzyme.coli DNA polymerase I gene (Klenow fragment) is cloned. coli cells in which the 3'-end two-thirds of the E. Klenow retains the polymerization fidelity of the holoenzyme without degrading 5' termini. Description: Klenow fragment, which requires template DNA and primer for its function, selectively catalyzes the transfer of dNTPs to the 3'-OH terminus of the primer that is complementary to the template.1)This enzyme is purified from E. The enzyme lacks the 5'→3' exonuclease activity of intact DNA polymerase I. Klenow Fragment is the large fragment of DNA Polymerase I that retains its 5'→3' polymerase, 3'→5' exonuclease and strand displacement activities.
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