T4 DNA Polymerase

$50.00$150.00


    125 Units - $50.00 (Cat.# 3221 )

    500 Units - $150.00 (Cat.# 3222 )

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

 Product SourceE.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.3221 3222

Additional information

Units

125 Units, 500 Units

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

You may also like…