DNA Polymerase I Large (Klenow) Fragment (500u)

DNA Polymerase I Large (Klenow) Fragment is a DNA-dependent DNA polymerase that lacks the 5´→3´ exonuclease activity of intact E. coli DNA Polymerase I but retains its 5´→3´ polymerase, 3´→5´ exonuclease and strand displacement activities. The enzyme is a 68kDa C-terminal fragment of DNA Polymerase I. 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. The 3´→5´ exonuclease activity can be used to generate blunt ends from a 3´-overhang.

Features - Benefits

• Flexible: DNA Polymerase I Large (Klenow) Fragment may be used in a variety of molecular applications. It is also active in many Promega 1X restriction enzyme buffers.
• May Be Heat-Inactivated: DNA Polymerase I Large (Klenow) Fragment is inactivated by heating at 75°C for 10 minutes.
• Provided with 10X Reaction Buffer: 500mM Tris-HCl (pH 7.2 at 25°C), 100mM MgSO₄, 1mM DTT.
• Choose Your Configuration: Learn more about our custom options for this product at: www.promega.com/custom/

Applications

• Filling-in or labeling 3´ ends of 5´ overhanging double-stranded DNA.
• Second-strand cDNA synthesis.
• DNA sequencing by the dideoxy method.

References

1. Anderson, S. et al. (1980) Nucl. Acids Res. 8, 1731–43.
2. Sanger, F. et al. (1977) Proc. Natl. Acad. Sci. USA 74, 5463–7.
3. Wallace, R.B. et al. (1980) Science 209, 1396–400.
4. Houdebine, L.M. (1976) Nucl. Acids Res. 3, 615–30.
5. Feinberg, A.P. and Vogelstein, B. (1983) Anal. Biochem. 132, 6–13.
6. Tabor, S. and Struhl, K. (1987) In: Current Protocols in Molecular Biology, Ausubel, F.M. et al., eds., John Wiley and Sons, New York, NY.
7. Joyce, C.M. and Grindley, N.D.F. (1983) Proc. Natl. Acad. Sci. USA 80, 1830–4.