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Taq-Purple DNA polymerase

Description

Taq-Purple DNA polymerase is an alternative to Taq DNA polymerase Unis (Cat. No. T037-T 039). In this product, concentration of Taq DNA polymerase is reduced to 1U/μl and storage buffer is supplemented with inert Dye for easy loading and control of the presence of the enzyme in PCR.

Inert red dye

  • The dye does not interfere with PCR and during electrophoresis in agarose gel it migrates faster than oligonucleotide primers. Therefore, the dye does not interfere with quantification of DNA amplicons.

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 takes < 1 min). Disadvantage is that it lacks a 3’->5’ exonuclease proofreading activity and this accounts for high error rate [about 1 error to 105 - 106 base pairs (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-Purple DNA polymerase is supplied at a  concentration 1 U/µl. Basic packaging is 1 tube with 500 U/500 µl (T107), 5 tubes with 500 U/500 µl (T108) or 10 tubes with 500 U/500 µl (T109).
  • Each tube of Taq Purple DNA polymerase is accompanied by 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: 20 mM Tris-HCl (pH 8.0 at 25oC), 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 0.5% Nonidet P-40, 0.5% Tween 20, inert red dye, 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 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 follow: 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 denatured cDNA, 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-Purple DNA polymerase is tested by SDS-PAGE. Enzyme migrates as a major band of 94 kDa. Material is nuclease free.
  • Each batch of Taq-Purple 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.

Downloads

Cat. No. Product name and specification Amount Price
T107 Taq-Purple DNA polymerase 500 U 91.25 EUR
T108 Taq-Purple DNA polymerase 5x 500 U 365 EUR
T109 Taq-Purple DNA polymerase 10x 500U 638.75 EUR
T107a Taq-Purple DNA polymerase + PCR dNTP mix 500 U + 100 µl 107.5 EUR
T108a Taq-Purple DNA polymerase + PCR dNTP mix 5x 500 U + 5x 100 µl 430 EUR
T109a Taq-Purple DNA polymerase + PCR dNTP mix 10x 500 U + 10x 100 µl 752.5 EUR
E101a PCR Enhancer together with 1000U of polymerases 1 ml 0 EUR