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DNA Polymerase I

Catalyzes the Template-Directed Polymerization of Nucleotides into Duplex DNA in a 5´→3´ Direction

  • Can be used in a variety of molecular applications
  • Is inactivated by heating at 68°C for 10 minutes
  • Supplied at a concentration of 5–10u/μl

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Description

Size

Size: 500u
Catalog number: M2051

Size: 2,500u
Catalog number: M2055

DNA Polymerase I catalyzes the template-directed polymerization of nucleotides into duplex DNA in a 5´→3´ direction. DNA Polymerase I possesses a 3´→5´ exonuclease activity or “proofreading” function, which lowers the error rate during DNA replication, and also contains a 5´→3´ exonuclease activity, which enables the enzyme to replace nucleotides in the growing strand of DNA by nick translation. The enzyme, purified from recombinant E. coli, is capable of catalyzing de novo synthesis of synthetic homopolymers and provides a convenient method for the preparation of a variety of defined DNA substrates. DNA Pol I is provided with 10X Reaction Buffer: 500mM Tris-HCl (pH 7.2 at 25°C), 100mM MgSO4, 1mM DTT.

Applications

  • Labeling of DNA to high specific radioactivity by nick translation.
  • Second-strand cDNA synthesis.

References

  1. Kelly, R.B. et al. (1970) J. Biol. Chem. 245, 39–45.
  2. Harwood, S.J. et al. (1970) J. Biol. Chem. 245, 5614–24.

Storage Buffer: 50mM Tris-HCl (pH 7.5 at 25°C), 1mM DTT, 0.1mM EDTA, 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 deoxyribonucleotides into TCA-insoluble material in 30 minutes at 37°C. The reaction conditions are: 67mM potassium phosphate (pH 7.4 at 25°C), 6.7mM MgCl2, 1mM DTT, 50μg/ml activated calf thymus DNA and 33μM dATP, dCTP, dGTP and dTTP (a mix of unlabeled and [3H]dTTP).

Protocols

Complete Protocol

DNA Polymerase I Protocol


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What’s in the box?
Size: 500u

Item Part # Size
DNA Polymerase 10X Buffer M195A 1 × 1ml
DNA Polymerase I M205A 1 × 500u

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

 

What’s in the box?
Size: 2,500u

Item Part # Size
DNA Polymerase 10X Buffer M195A 5 × 1ml
DNA Polymerase I M205B 1 × 2,500u

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB