| Catalyzes 5’=>3’ synthesis of DNA. Enzyme has 3’=>5’ exonuclease activity but lacks 5’=>3’ exonuclease activity. T4 DNA polymerase possessing 3'=>5' exonuclease activity exhibits greater activity on single-stranded DNA than on double-stranded DNA. |
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| Catalog # | D3943 |
| Source | E. coli with a cloned gene43 of bacteriophage T4. |
| Applications | • Blunting of DNA ends: fill-in 5'-overhangs or/and removal of 3'-overhangs, see protocol |
| • Blunting of PCR products with 3’-dA overhangs. |
| • Synthesis of labeled DNA probes by the replacement reaction. |
| • Oligonucleotide-directed site-specific mutagenesis. |
| • Ligation-independent cloning of PCR products. |
| 5X Reaction Buffer | D3943-05: DNA Polymerase T4 Buffer, 5X: Supplied as a liquid in 335mM Tris-HCl, pH 8.8 at 25°C, 33mM magnesium chloride, 5mM DTT, 84mM (NH4)2SO4. |
| Unit Definition | One unit of T4 DNA Polymerase catalyzes the incorporation of 10nmoles of deoxyribonucleotides into a polynucleotide fraction (adsorbed on DE-81) in 30min at 37°C. |
| Activity Assay | Assayed in the following mixture: 67mM Tris-HCl, pH 8.8, 6.7mM MgCl2, 1mM DTT, 16.7mM (NH4)2SO4, 0.2mg/ml BSA, 0.033mM of each dNTP, 0.4meq/ml [3H]-dTTP, 0.2mM heat-denatured and nuclease-digested calf thymus DNA. |
| Quality Control | No conversion of covalently closed circular DNA to nicked DNA was detected after incubation of 10 units of T4 DNA Polymerase with 1ug of pUC19 DNA in 50ul reaction buffer for 4 hours at 37°C. |
| Storage and Stability | Aliquot to avoid repeated freezing and thawing.. Store at -20°C. Aliquots are stable for 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. |
| Molecular Weight | ~104kD |
| Concentration | ~5u/ul |
| Form | Supplied as a liquid in 20mM potassium phosphate, pH 7.5, 200mM potassium chloride, 2mM DTT, 50% glycerol. |
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| Important Note | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
