Taq DNA polymerase
Taq DNA polymerase is a thermostable enzyme isolated from Thermus aquaticus. The enzyme catalyzes synthesis of complementary DNA strand in the 5’->3’ direction and also possesses a 5’->3’ exonuclease activity. During amplification of DNA fragments, Taq polymerase adds at 3’ end an adenosine overhang. This can be utilized for cloning of PCR-generated DNA fragments. Advantage of the enzyme is its high processivity [amplification of 1000 base pairs (bps) takes < 1 min]. Disadvantage of the enzyme is that it lacks a 3’->5’ exonuclease proofreading activity and this accounts for high error rate (about 1 error to 105 - 106 base bps). The major usage of the enzyme is in diagnostic analysis for amplification of DNA fragments up to 5000 bps.
| Cat.no. | Name | Package | Price | |
|---|---|---|---|---|
| T032 | Taq DNA polymerase | 500 U | 2 189,00 Kč | |
| T033 | Taq DNA polymerase | 5x 500 U | 8 756,00 Kč | |
| T034 | Taq DNA polymerase | 10x 500 U | 15 323,00 Kč | |
| T034a | Taq DNA polymerase + PCR dNTP mix | 10x 500 U + 10x 100 µl | 18 326,00 Kč | |
| T032a | Taq DNA polymerase + PCR dNTP mix | 500 U + 100 µl | 2 618,00 Kč | |
| T033a | Taq DNA polymerase + PCR dNTP mix | 5x 500 U + 5x 100 µl | 10 472,00 Kč |
Product description
Taq DNA polymerase is a thermostable enzyme isolated from Thermus aquaticus. The enzyme catalyzes synthesis of complementary DNA strand in the 5’->3’ direction and also possesses a 5’->3’ exonuclease activity. During amplification of DNA fragments, Taq polymerase adds at 3’ end an adenosine overhang. This can be utilized for cloning of PCR-generated DNA fragments. Advantage of the enzyme is its high processivity [amplification of 1000 base pairs (bps) takes < 1 min]. Disadvantage of the enzyme is that it lacks a 3’->5’ exonuclease proofreading activity and this accounts for high error rate (about 1 error to 105 - 106 base bps). The major usage of the enzyme is in diagnostic analysis for amplification of DNA fragments up to 5000 bps.
Technical data
Components and packaging
- Taq DNA polymerase is supplied at a concentration 5 U/µl. Basic packaging is 1 tube with 500 U/100 µl (T032), 5 tubes with 500 U/100 µl (T033) or 10 tubes with 500 U/100 µl (T034).
- Each tube of Taq DNA polymerase is accompanied with a tube with 10x concentrated react buffer containing MgCl2 (1.5 ml). If different concentration of MgCl2 is required, a tube with 10x concentrated reaction buffer without MgCl2 (1.5 ml) and a tube with 25 mM MgCl2 (0.5 ml) should be ordered (Cat. No. T035).
Storage
- At temperature -20oC ± 5°C. Material can be repeatedly defrosted.
Composition
- Storage buffer for Taq DNA polymerase: 20 mM HEPES (pH 7.9 at 25oC), 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 0.5 mM PMSF, stabilizers, 50% glycerol.
- 10x reaction buffer: 100 mM Tris-HCl, pH 8.8 (at 25oC), 500 mM KCl, 1% Triton X-100, 15 mM MgCl2.
Activity
- One unit of Taq DNA polymerase is defined as the amount of enzyme, which catalyzes incorporation of 10 nmol dNTPs within 30 min at 72oC into trichloracetic acid precipitable material. Reaction conditions are as follows: 10 mM Tris-HCl (pH 8.8 at 25oC), 50 mM KCl, 0.1% Triton X-100, 1.5 mM MgCl2, 200 µM dATP, dCTP, dGTP and [α-32P]dTTP, 50 µg/ml activated salmon testes DNA, 0.5 µM primer and 0.2 – 0.5 U of enzyme in the volume of 50 µl.
Purity and quality control
- Purity of Taq DNA polymerase is tested by SDS-PAGE. After staining with Coomassie blue, enzyme migrates as the major band of 94 kDa. Material is nuclease free.
- Each batch of Taq DNA polymerase is tested for its ability to amplify DNA fragment from mammalian genomic DNA by PCR. The results are verified by electrophoresis in agarose gel in the presence of ethidium bromide; only DNA band of the expected size is present.