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DNA Polymerase I (2,500u)

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M2055

Description

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.

Features - Benefits

  • Flexible: DNA Polymerase I may be used in a variety of molecular applications.
  • May Be Heat-Inactivated: DNA Polymerase I is inactivated by heating at 68°C for 10 minutes.
  • 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.

Specifications

QC Tests

Activity, SDS-PAGE/purity, endonuclease.

Source

Recombinant E. coli strain.

Storage Buffer

50mM Tris-HCl (pH 7.5 at 25°C), 1mM DTT, 0.1mM EDTA, 50% (v/v) glycerol.

Storage Conditions

Store at –20°C.

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).

For product intended use please see Patents & Disclaimers tab.

Protocols and manuals

Related citations

Resources

Use Restrictions

M2051, M2055 For Research Use Only. Not for Use in Diagnostic Procedures.

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