| Description |
T4 DNA Polymerase catalyzes the extension of a primed DNA template (in the presence of template and primer) in the 5′→ 3′ direction. This enzyme exhibits a powerful 3′→ 5′ exonuclease activity which is much more active than that found in DNA Polymerase I (E. coli), while lacking any inherent 5′→ 3′ exonuclease or strand displacement functions.
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| Applications |
- Blunting of DNA ends: fill-in of 5'-overhangs or/and removal of 3'-overhangs
- Blunting of PCR products with 3'-dA overhangs
- Single strand deletion subcloning (Ligation-independent cloning of PCR products).
- Second strand synthesis in site-directed mutagenesis.
- Probe labeling using replacement synthesis.
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| Source |
A recombinant E. coli strain carrying a cloned gene 43 of bacteriophage T4. |
| Unit Definition |
One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid-precipitable material in 30 minutes at 37°C. |
| Components |
- T4 DNA Polymerase: 3,000 U/ml in 100 mM KPO4, 1.0 mM DTT, 0.1 mM EDTA, 50% glycerol, pH 6.5 @ 25°C
- 10X Reaction Buffer: 500 mM NaCl, 100 mM Tris-HCl, 100 mM MgCl2, 10 mM DTT, pH 7.9 @ 25°C
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| Quality Control |
The absence of endodeoxyribonucleases confirmed by appropriate quality tests. |
| Storage Condition |
-20 °C
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