![]() Increase in the apparent rate of primer elongation from 8 nt/s (with excess polymerase) to an average rate of 30 nt/s at 30˚C was also observed. 6B)." P.31196 left column 4th paragraph: "Analysis of extension products from single interactions of the enzyme with the M13 template revealed a >200-fold increase in enzyme processivity when the concentration of MgCl2 was reduced from 8 to 1 mM. ![]() The extension efficiency was also affected by the up-stream overhanging structure of the primertemplate complex. The enzyme exhibits DNA synthesis and proofreading (35) nuclease activities, and, in the absence of the holoenzyme’s (53) nuclease domain, displays a moderate strand displacement activity during DNA synthesis. 2, time course) to ≥30 nt/s (RFII in 4 min, Fig. Under the condition we conducted, the shortest primers polymerized by Klenow fragment (KF) and Taq DNA polymerase in our experiments were respectively heptamer and octamer. Klenow Fragment is a mesophilic DNA polymerase derived from the E.coli Polymerase I DNA-dependent repair enzyme. However, polymerization behavior of short primers in the primer extension process has not been systematically explored. The apparent elongation rate during RFII formation from 8 nt/s (RFII in 15 min, Fig. Polymerization behavior of Klenow fragment and Taq DNA polymerase in short primer extension reactions DNA polymerases amplify DNA fragments through primer extension reactions. Inclusion of the Escherichia coli helix destabilizing protein allowed the viral enzyme, which lacks strand displacement activity, to utilize a singly primed M13 DNA template."Ībstract: "A primer extension rate of 30 nt/s was observed, and > or = 2000 nt were added per binding event." P.31194 right column bottom paragraph: "Indeed, reducing the MgCl2 concentration from 8 to 1 mM increased PCR (ordinary and high-throughput) Primer Extension Microarray Analysis Denaturing high performance liquid chromatography (DHPLC). report here an investigation of the intrinsic processivity of this enzyme on both natural and homopolymeric DNA templates. Synthesis of probes by random primers labeling method (2). Applications: Fill-in of 5 overhangs (1). adding a radio-deoxynucleotide to the 3-end in a templated extension reaction. DNA Polymerase I, Large (Klenow) Fragment is a DNA polymerase enzyme that lacks the 5' to 3' exonuclease activity of intact DNA Polymerase I, but does exhibit the 5' to 3' DNA polymerase and 3' to 5' exonuclease activities. abstract, p.31194 right column bottom paragraph & p.31196 left column 4th paragraph PubMed ID 7983061Ībstract: "The polymerization and proofreading activities of the vaccinia virus DNA polymerase reside within a 116-kDa catalytic polypeptide. In this recombinant DNA work, the appropriate Klenow fragment gene was. ![]() In vitro analysis of parameters affecting processivity. Primer extension rate by DNA polymerase Value ![]()
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