T4 DNA Polymerase
B buffer for 100% activity FastDigest buffer for 100% activity G buffer for 100% activity O buffer for 100% activity Recombinant enzyme R buffer for 100% activity Tango buffer for 100% activity Thermal inactivation at 75°C in 10 min
T4 DNA Polymerase
T4 DNA Polymerase is template-dependent and catalyzes the 5'-3' synthesis from primed single-stranded DNA.
Thermo ScientificT4 DNA Polymerase, a template-dependent DNA polymerase, catalyzes 5'-3' synthesis from primed single-stranded DNA. The enzyme has a 3'-5' exonuclease activity, but lacks 5'-3' exonuclease activity.
- Stronger 3'-5' exonuclease activity on single-stranded than on double-stranded DNA and greater (more than 200 times) than DNA polymerase I, E. coli,and Klenow fragment (see Reference1)
- Active in Thermo Scientific restriction enzyme, PCR, RT and T4 DNA Ligase buffers
- Blunting of DNA ends: fill-in of 5'-overhangs or/and removal of 3'-overhangs(see References1, 2)
- Blunting of PCR products with 3'-dA overhangs
- Synthesis of labeled DNA probes by the replacement reaction(see Reference3)
- Oligonucleotide-directed site-specific mutagenesis(see Reference4)
- Ligation-independent cloning of PCR products(see References5, 6)
5× Reaction Buffer 335mM Tris-HCl (pH8.8 at 25°C), 33mM MgCl2, 5mM DTT, 84mM (NH4)2SO4 Concentration 5U/µL Definition of Activity Unit
- One unit of the enzyme catalyzes the incorporation of 10 nmol of deoxyribonucleotides into a polynucleotide fraction (adsorbed on DE-81) in 30min at 37°C.
- Enzyme activity is 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, 0.4 MBq/mL [3H]-dTTP, and 0.2mM heat-denatured and nuclease-digested calf thymus DNA.
Hazardous No Inactivation Inactivated by heating at 75°C for 10min. Inhibition Inhibitors: metal chelators, nucleotide analogs 2(p-n-butylanilino)-dATP, N2-(p-n-butylphenyl)-dGTP), SH-blocking compounds (see Reference 7) Molecular Weight 104kDa monomer Quality Control The absence of endodeoxyribonucleases confirmed by appropriate quality tests. Source E.coli cells with a cloned gene 43 of bacteriophage T4 Storage Buffer The enzyme is supplied in 20mM potassium phosphate (pH7.5), 200mM KCl, 2mM DTT, and 50%(v/v) glycerol. Storage Condition -20 C
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