Application Example

Figure: Sensitive HotStart PCR amplification with Perpetual Taq DNA Polymerase (Cat. No. E2700) in a size range between ~1 and 15 kb. Molecular weight marker M1 (Perfect™ 1kb DNA Ladder). Lanes 1.1 to 15 kb: PCR amplification reactions (each reaction 1.25 U EURx Perpetual Taq DNA Polymerase, Pol Buffer B, 0.2 mM dNTPs; 50 µl reaction volume).

Characteristics

• HotStart Taq: For PCR applications requiring "HotStart", such as sensitive fingerprinting techniques or optical PCR methods.
• High Quality Antibody: Contains a top quality, highly specific, monoclonal anti-Taq antibody with one single, precisely defined denaturation temperature. And, yes, we know, high quality anti-Taq antibodies are rare, extremely rare. And we are aware that it will be hard trying to find a better antibody anywhere else.
• Optimal HotStart Technology: Initial inhibition of Taq DNA polymerase mediated by high quality anti-Taq antibodies is regarded as the gold standard in HotStart PCR technology. In many, if not most scenarios, carefully prepared, anti-Taq antibody-inhibited Taq DNA polymerase preparations, such as Perpetual Taq, provide superior results - more precise, much cleaner and more reliable with regards to reproducibility -, as compared to chemically or nucleic acid (aptamer) - inhibited HotStart DNA polymerase formulations. Therefore, Perpetual Taq DNA polymerase is recommended for PCRs aiming at highest possible quality, e.g. for diagnostic PCRs.
• No Primer Dimers: Anti-Taq antibody prevents formation of primer-dimers during the first heating step from RT to 95°C. This step is probably the most sensitive step throughout the whole PCR process, since primer dimers are amplified preferentially due to their short size and - of course - due to their perfectly matching primer sequences.
• All Buffers Inclusive: Ships complete with non-colored plus with colored buffers for direct gel loading.
• Thorough and Gentle Enzyme Purification: High purity and efficient enzymatic activity as a result of thorough but gentle enzyme purification.
• Optimized Towards Quality:
  • No false positives: No detection of PCR-amplificable DNA, as tested in 40+ cycle PCR reactions with various amplification targets of human, eukaryotic, bacterial and viral origin (including bacterial 16S rRNA genes and human viral genes).
  • Extreme Purity: PAGE gel analysis of purified enzyme and of purified antibody, respcectively, reveal presence of just one single band, as one would expect for a >95% pure preparation.
  • Devoid of Nucleases and Proteases: Sensitive enzymatic tests reveal absence of any contaminating nucleases or proteases.
  • Strict Quality Control: Optimized towards high enzyme quality: Both, Taq DNA polymerase and anti-Taq antibody, respectively, are continuously and extensively monitored. Strict quality controls ensure continuous reproducible high purity and performance of the HotStart enzyme preparation, as opposed to certain infamous "crude extracts with intrinsic DNA polymerase activity" offered by some third parties.
  • Precise Conformity with Unit Specification: Strict and precise adjustment of enzymatic activity towards the regular, commonly accepted unit definition. One unit of enzyme is exactly one unit - and not less. At 74°C, one unit catalyzes the incorporation of precisely 10 nmoles of dNTP into acid-insoluble material within 30 min, no more, no less.
  • Recombinant Taq: Optimized, improved performance as compared to → Native Taq DNA Polymerase [Cat. No. E2504].
• Good Value for Money: At this price, this high quality enzyme preparation is truly a bargain.
• Enzyme Properties: Intrinsic exonuclease activities: 5'-3' present, 3'-5' none. No proofreading activity.
• Cloning: Enables both TA- and blunt end cloning.
• Also available as: → Perpetual Taq PCR Master Mix [Cat. No. E2740].

Perpetual Taq DNA Polymerase - Package Contents

• Perpetual Taq DNA Polymerase
• 10x Buffer Pol B (Standard reaction buffer containing MgCl₂)
• 10x Buffer Pol C (Purple coloured reaction buffer, for direct gel loading)
• 10x Buffer Pol A (Optimization reaction buffer without MgCl₂)
• 25 mM MgCl₂ solution

Perpetual Taq DNA Polymerase - Kit Package Contents

• Same as Perpetual Taq DNA Polymerase, plus ultrapure dNTP solution

Additional Resources

 PCR Logsheet