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Taq DNA polymerase
Features: Concentrated up to 50 000 U/ml – by request.
Description: Purified from E.coli strain carrying plasmid with the cloned gene encoding Thermus aquaticus DNA polymerase. Taq DNA polymerase catalyses 5’→3’ synthesis of DNA. The enzyme has no detectable 3’→5’ proofreading exonuclease activity, but possesses 5’→3’ exonuclease activity.
Unit Definition: One unit of enzyme catalyses incorporation of 10 nanomoles of deoxyribonucleotides into acid-insoluble polynucleotide fraction in 30 min at 70°C.
Activity assay: 50 mM Tris-HCl (pH 8.0 at 25°C), 50 mM NaCl, 10 mM MgCl2, 200 μM dATP, 200 μM dCTP, 200 μM dGTP, 50 μM [3H] dTTP, 0,25 mg/ml activated calf thymus DNA.
Storage conditions: -20° C in 50 mM Tris-HCl (pH 8.0 at 25°), 50 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 50% glycerol and 1% triton X-100.
Unit Definition: One unit of enzyme catalyses incorporation of 10 nanomoles of deoxyribonucleotides into acid-insoluble polynucleotide fraction in 30 min at 70°C.
Activity assay: 50 mM Tris-HCl (pH 8.0 at 25°C), 50 mM NaCl, 10 mM MgCl2, 200 μM dATP, 200 μM dCTP, 200 μM dGTP, 50 μM [3H] dTTP, 0,25 mg/ml activated calf thymus DNA.
Storage conditions: -20° C in 50 mM Tris-HCl (pH 8.0 at 25°), 50 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 50% glycerol and 1% triton X-100.
Quality control: Endo-, exodeoxyribonucleases, ribonucleases free.
Applications:- Amplifications of DNA fragments by polymerase chain reaction (PCR).
- DNA labelling with radionucleotides, digoxygenin or biotin.
- DNA sequencing.