| Description: |
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DFS (DNA Free Sensitive) Taq DNA Polymerase is a thermostable enzyme of approximately 94kDa isolated from eubacterium Thermus aquaticus strain YT-1(1). This unmodified enzyme replicates DNA at 72°C. The enzyme catalyzes the polymerization of nucleotides into duplex DNA in the 5´-->3´ direction in the presence of magnesium ions and possesses a 5´-->3´ exonuclease activity. The enzyme is highly purified and is free of nonspecific endo- or exonucleases. Taq DNA polymerase leaves single 3'-dA nucleotide overhangs on their reaction products.
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| Performance and purity tests: |
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Taq DNA polymerase effectively directs PCR with the template up to 5 kb in length. Enzyme was tested on the absence of endonuclease and nickase activities. No traces of bacterial DNA were detected in PCR reaction with “no template” test with the primers complementary to the conservative region of 16S ribosomal gene.
The following tests are performed with each lot of DFS Taq DNA polymerase: |
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- PCR with various templates – human and bovine genomic DNA, Phage Lambda DNA;
- Exo-endo nucleases contamination tests;
- “no primers” test with Lambda DNA cycling without primers;
- “no template” test with the primers complementary to the conservative region of 16S bacterial ribosomal genes;
- storage (3 days at room temperature) test – no change in performance.
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| Applications |
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DNA-free Taq DNA polymerase is suitable for all regular applications – PCR, primer extension reactions etc. DFS Taq DNA polymerase is free from bacterial DNA and it is especially recommended for the work with bacterial DNA.
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| Sensitivity |
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of PCR reaction with DFS Taq DNA polymerase in the optimal conditions is very high – in some reactions less than 6 DNA molecules were detected. Enzyme has a very good performance in single-copy gene PCR from genomic mammalian DNA.
In contrast to Bioron enzyme, Taq DNA polymerases from the variety of uppliers contain contaminating DNA, with DNA-contaminated Taq DNA polymerase one should be ready to observe false-positive PCR results in some cases. |
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| Concentration |
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5000 units/ml |
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| Storage buffer |
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10 mM K-phosphate, pH 7.4, 0.1 mM EDTA, 50% glycerol, 0.1% Triton X-100, 0.1% Tween 20. |
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| Unit definition |
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One unit of activity is the amount of enzyme required to incorporate 10 nmoles of dNTP into acid-insoluble DNA fraction in 30 minutes at 72℃. |
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| Reaction buffer (x10)” incomplete”: |
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160 mM (NH4)2SO4, 670mM TrisHCl pH8,8, 0,1% Tween-20 |
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| Reaction buffer (x10) “complete”: |
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160 mM (NH4)2SO4, 670mM TrisHCl pH8,8, 0,1% Tween-20, 25mM MgCl2 |
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| Reaction buffer (x10) “complete II KCl”: |
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500 mM KCl, 100mM TrisHCl pH8,8, 0,1% Tween-20, 15mM MgCl2 |
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| Additionally provided: |
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1 Tube MgCl2 (100 mM) |
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| Recommended MgCl2 concentration |
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1,5mM - 6mM |
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| Storage conditions |
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Recommended storage temperature is -20℃, meanwhile, the enzyme is stable at room temperature at least for 3 days without any loss of activity |
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| Reference: |
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1.Kaledin, A.S., Sliusarenko, A.G. and Gorodetskii, S.I. (1980) Biokhimiya 45, 644(Rus) |
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Catalog #
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conc.
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Pack size
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101005
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5,0u/l
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500 u
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101025
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5,0u/l
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2500 u
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101100
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5,0u/l
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10000 u
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101500
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5,0u/l
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50000 u
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