5 PRIME Hot Master Taq DNA Polymerase

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5 PRIME Hot Master Taq DNA Polymerase: –
Innovative Hot-Start/Cold-Stop technology for highly specific
Hot-Start PCR without enzyme activation.

Contains TaqMaster PCR Enhancer that provides robust amplification of
contaminated and difficult samples.

Key Benefits:

  • Optimal results with highly specific amplification
    • Hot-start is a well established method for improving PCR specificity.  5 PRIME Hot Master Taq DNA polymerase is designed to reduce or eliminate any non-specific products that result from mispriming during PCR.  Conventional hot-start technologies, such as antibody-mediated inhibition or chemical blocking of DNA polymerases, have limitations, such as long initial activation steps that can reduce the performance of the DNA polymerase and compromise specificity control.
      To avoid this disadvantages, 5 PRIME Hot Master Taq DNA Polymerase features a superior hot-start PCR technology: a temperature dependent inhibitory ligand completely inhibits Taq polymerase activity at low temperatures.
      At high temperatures, the inhibitor is released and full Taq activity is immediately restored. Since the process is reversible, the “cold stop” aspect of the inhibitor has the potential to block enzyme activity in every low-temperature cycle of the PCR, ensuring optimal results. Unlike standard Taq polymerases, HotMaster Taq reactions can be
      set up at room temperature.
  • Savings of time and effort by optimized PCR conditions
    • The 5 PRIME HotMaster Taq DNA Polymerase is provided with a 10x self-adjusting Mg buffer
      The formulation adjusts the Mg 2+ concentration automatically, eliminating the need for optimizing this critical component.
  • Excellent for fast PCR protocols – no enzyme activation
    • The initial heat-activation step required by  standard hot-start PCR enzymes prior to cycling, typically up to 15 minute incubation at 95°C, can reduce enzyme activity and result in lower yield of PCR product. HotMaster Taq does not require an initial heat-activation step and can efficiently amplify short fragments as well as fragments up to several kb (e.g. up to 5 kb from genomic DNA) with high specificity. This combination of high efficiency and high specificity guarantees detection of extremely low levels of target DNA. Even the presence of high levels of non-template DNA, HotMaster Taq can amplify less than 10 target DNA molecules.
The 5 PRIME HotMaster Taq DNA Polymerase uses an innovative technology to achieve Hot Start PCR. A thermostable inhibitor (patent-pending) of the Taq DNA Polymerase is released at high temperatures, thereby immediately activating the enzyme. The HotMaster technology not only provides Hot Start control at reaction setup, but also Cold Stop during the annealing step of each and every cycle of PCR. The result is a unique Hot Start/Cold Stop technology.
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Fast PCR Amplification of a 131 bp fragment of the human TFN gene with standard Taq, HotMaster Taq and conventional Hot Start enzymes. Used protocol: 1 sec 95 °C denaturation, 1 sec 55 °C annealing, 5 sec 72 °C extension. Initial denaturation was 2 min at 95 °C prior to PCR or 10 min for the chemically modified enzyme respectively.

 

Packaging:

  • HotMaster Taq DNA Polymerase from 5 PRIME is packaged with 10x Reaction buffer.

Storage & Dilution Buffer:

  • 25 mM Tris-HCl pH 8.0, 35 mM KCl, 0.1 mM EDTA, 1 mM DTT, 50% glycerol, 0.5% Tween 20, 0.5% IGEPAL, CA-630 and stabilizers.

Reaction Buffer:

  • 10x HotMaster Taq Buffer with 25 mM Mg2+

Unit Definition:

  • One unit is defined as the amount of enzyme that incorporates 10 nmoles of dNTPs into acid-insoluble form within 30 min at 74°C under the assay conditions, using reaction conditions: 25 mM TAPS (N-tris-(hydroxymethyl)-methyl-3-amino-propanesulfonic acid, sodium salt) pH 9.3 (at 25°C); 50 mM KCI; 2 mM MgCI2; 1 mM b-mercaptoethanol; 200 µmol each dATP, dGTP, dTTP; 100 µM dCTP (a mix of unlabeled and [a-32P]-labeled); 12.5 µg activated salmon sperm DNA in a final volume of 50 µl.

Associated Activities:

  • No endo- or exonuclease activity was detected after a 14–16 hour incubation at 37°C of the polymerase with lambda/Hind III fragments and purified plasmid DNA.
Source:
  • Recombinant and modified form of the Thermus aquaticus YT1 1 DNA-Polymerase-Gene expressed in E. coli DH 1. (1) Kaledin, A.S. et al. (1980) Biokhimiya 45, 494)

Concentration:

  • 5 units/µl

Storage Requirements:

  • Store at –20°C in a constant temperature freezer.

Ordering:

  • Cat # 2200300: 5 PRIME Hot Master Taq DNA Polymerase – 100U
  • Cat # 2200320: 5 PRIMEHot Master Taq DNA Polymerase – 1000U
  • Cat # 2200330: 5 PRIMEHot Master Taq DNA Polymerase – 5000U

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