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DNA Polymerase I Large (Klenow) Fragment

DNA-Dependent DNA Polymerase that Lacks the 5´→3´ Exonuclease Activity

  • Use in a variety of molecular applications
  • Heat-inactivate DNA Polymerase I Large (Klenow) Fragment by heating at 75°C for 10 minutes
  • Is active in many Promega 1X restriction enzyme buffers

Size

Catalog number selected: M2201

$ 114.00 Your price: Log In

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DNA Polymerase I Large (Klenow) Fragment
150u
$ 114.00
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DNA Polymerase I Large (Klenow) Fragment is a 68kDa C-terminal fragment of E. 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. 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. The DNA-dependent DNA polymerase is 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/

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.

Storage Buffer: 50mM Tris-HCl (pH 7.5), 1mM DTT, 0.1mM EDTA and 50% (v/v) glycerol.

Source: Recombinant E. coli strain.

QC Tests: Activity, SDS-PAGE/purity, endonuclease.

Unit Definition: One unit is defined as the amount of enzyme that incorporates 10nmol of total deoxyribonucleotides into TCA-insoluble material in 30 minutes at 37°C. The reaction conditions are: 67mM potassium phosphate (pH 7.5 at 25°C), 6.7mM MgCl2, 1mM DTT, 50μg/ml activated calf thymus DNA and 33μM each of dATP, dCTP, dGTP and dTTP (a mix of unlabeled and [3H]dTTP).

Protocols

Specifications

You are viewing: 150u Change Configuration

What's in the box?

Item Part # Size

DNA Polymerase 10X Buffer

M195A 1 x 1ml

DNA Polymerase I Large (Klenow) Fragment

M220A 1 x 150u

SDS

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Certificate of Analysis

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Use Restrictions

For Laboratory Use. Outside of the United States, this product is intended for research use only unless otherwise stated.

Storage Conditions

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Specifications

You are viewing: 500u Change Configuration

What's in the box?

Item Part # Size

DNA Polymerase 10X Buffer

M195A 1 x 1ml

DNA Polymerase I Large (Klenow) Fragment

M220C 1 x 500u

SDS

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Certificate of Analysis

Search for Specific Certificate:

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Use Restrictions

For Laboratory Use. Outside of the United States, this product is intended for research use only unless otherwise stated.

Storage Conditions

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