Klenow Fragment, Exonuclease Minus (100u)

DNA Polymerase I Large (Klenow) Fragment, Exonuclease Minus, is a DNA-dependent DNA polymerase that lacks both the 5´→3´ and the 3´→5´ exonuclease activities present in intact E. coli DNA Polymerase I. It is used for random primer labeling and in strand displacement amplification. Klenow Fragment, Exonuclease Minus, will leave a single-base 3´ overhang on a significant proportion of DNA fragments during fill-in of 5´-overhangs. Therefore, this enzyme is not recommended for preparation of blunt-ended fragments for ligation.

Features - Benefits

• Provided with 10X Reaction Buffer: 500mM Tris-HCl (pH 7.2 at 25°C), 100mM MgSO₄, 1mM DTT.
• May Be Heat-Inactivated: DNA Polymerase I Large (Klenow) Fragment, Exonuclease Minus, is inactivated by heating at 75°C for 10 minutes.

Applications

• Random primer labeling.
• Strand Displacement Amplification (SDA).
• DNA sequencing by the dideoxy method.

References

1. Derbyshire, V. et al. (1988) Science 240, 199–201.
2. Sanger, F. et al. (1977) Proc. Natl. Acad. Sci. USA 74, 5463–7.
3. Feinberg, A.P. and Vogelstein, B. (1984) Anal. Biochem. 137, 266–7.
4. Walker, G.T. (1993) PCR Meth. Appl. 3, 1–6.
5. Clark, J.M. et al. (1987) J. Mol. Biol. 198, 123–7.