(e) Calculated polymerisation rate of the Klenow fragment. (1987) In: Current Protocols in Molecular Biology, Ausubel, F.M. Three substrates with a 20-, 40- and 60- and overhang with a total 80 bases were incubated with 1 nM Klenow fragment. Ĭhoose Your Configuration: Learn more about our custom options for this product at: References The DNA-dependent DNA polymerase is provided with 10X Reaction Buffer. The 3´→5´ exonuclease activity can be used to generate blunt ends from a 3´-overhang. Pol I can be cleaved by mild treatment with subtilisin into two fragments the larger fragment is known as the Klenow fragment. The 5´→3´ polymerase activity of Klenow Fragment can be used to fill in 5´-protruding ends with unlabeled or labeled dNTPs, to sequence single- or double-stranded DNA templates, for in vitro mutagenesis using synthetic oligonucleotides, for cDNA second-strand synthesis and to generate single-stranded DNA probes. Escherichia coliDNA Pol I can carry out three enzymatic reactions: It possesses 5' 3' DNA polymerase activity and 3' 5' and 5' 3' exonuclease activity. coli DNA Polymerase I that lacks the 5´→3´ exonuclease activity of intact DNA polymerase I but retains its 5´→3´ polymerase, 3´→5´ exonuclease and strand displacement activities.

coli DNA Polymerase I that has lost the 5-3 exonuclease activity while retaining the original 5-3. DNA Polymerase I Large (Klenow) Fragment is a 68kDa C-terminal fragment of E. Can we run PCR reaction using Klenow fragment (DNA polymerase without exonuclease activity) 2. The Klenow fragment is a truncated version of the E.