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T4 DNA Polymerase

$50.00$150.00


    125 Units - $50.00 (Cat.# 3221 )

    500 Units - $150.00 (Cat.# 3222 )

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Description

Intact Genomics T4 DNA Polymerase has both a DNA-dependent DNA polymerase activity and a potent 3´→5´ exonuclease activity.  

The purity of this enzyme is > 95% homogeneity as determined by SDS-PAGE using Coomassie® Blue staining (see figure below).

Picture

Additional information

Units

125 Units, 500 Units

Product Source
E.coli cells with a cloned gene of bacteriophage T4 DNA Polymerase.

Applications

  • 3´-overhang removal to form blunt ends (1, 2).
  • 5´-overhang fill-in to form blunt ends (1, 2).
  • Probe labeling using replacement synthesis (2).
  • DNA library preparation for Next-generation sequencing.
  • Ligation-independent cloning of PCR products.
  • Second strand synthesis in site-directed mutagenesis (3).

Product Includes

  1. T4 DNA Polymerase
  2. 10x T4 DNA Polymerase Buffer

Storage Temperature
–20 °C

Storage Buffer
50 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA,
50% Glycerol, pH 7.5 @ 25 ºC

10X T4 DNA Polymerase Reaction Buffer

500 mM Tris-HCl, 100 mM MgCl2, 50 mM dithiothreitol, pH 7.5 @ 25°C.

Unit Definition

One unit of T4 DNA Polymerase converts 10 nmol of dNTPs into acid-insoluble material in 30 minutes at 37°C under standard assay conditions.

Inactivation

Inactivated by heating at 70°C for 15 min.

Quality Control Assays

T4 DNA Polymerase is free from detectable endonuclease and RNase activities.

Blunting ends by 3’ overhang removal or 3’ recessed end fill-in:

  1. Dissolve DNA in 1x reaction buffer supplemented with 100 μM dNTPs. 
  2. Add 1 unit T4 DNA Polymerase per μg DNA.
  3. Incubate at room temperature for 5-30 minutes. Stop reaction by heating at 70°C for 20 minutes.

References

  1. Tabor, S. and Struhl, K. (1989). DNA-Dependent DNA Polymerases. Current Protocols in Molecular Biology. 3.5.10-3.5.12. New York: John Wiley & Sons, Inc.
  2. Sambrook, J. et al. (1989). Molecular Cloning: A Laboratory Manual. (2nd ed.), 5.44-5.47. Cold Spring Harbor: Cold Spring Harbor Laboratory Press.
  3. Kunkel, T.A. et al. (1987). Methods Enzymol. 154, 367-382.

T4 DNA Polymerase Manual

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