KAPATaq HotStart

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KAPATaq HotStart DNA Polymerase is based on the single- subunit, wild-type Taq DNA polymerase of the thermophilic bacterium Thermus aquaticus. In the HotStart formulation, the enzyme (which is purified from recombinant E. coli) is combined with a proprietary antibody that inactivates the enzyme until the first denaturation step. This eliminates spurious amplification products resulting from non-specific priming events during reaction setup and initiation, and increases overall reaction efficiency and sensitivity. Antibody-based hotstart PCR enzymes offer superior performance compared to hotstart formulations based on chemical modification of the enzyme, as enzyme re-activation is faster, more complete and is optimal at pH values which are also optimal for polymerase activity.

KAPATaq HotStart Buffer is a novel buffer designed for optimal enzyme re-activation and performance. The proprietary formulation also facilitates specific primer annealing, which translates to higher yields of specific product when compared to traditional Taq buffers. The 5x buffer is supplied without MgCl2 for optimal flexibility. KAPATaq HotStart DNA Polymerase may, however, be used in combination with any standard Taq buffer with a pH of 8.3 or higher.

KAPATaq HotStart DNA Polymerase has 5’→3’ polymerase and 5’→3’ exonuclease activities, but no 3’→5’ exonuclease (proofreading) activity. The enzyme has an error rate of approximately 1 error per 2.2 x 105 nucleotides incorporated. PCR products generated with KAPATaq HotStart are A-tailed and may be cloned into TA cloning vectors.

Product Applications

KAPATaq HotStart DNA Polymerase is ideally suited for:

  • High throughput PCR
  • Amplification of low copy DNA templates
  • Multiplex PCR
  • Specific amplification of complex templates
  • RT-PCR

KAPATaq HotStart outperforms the competition

A 500 bp fragment of the CCR5 gene was amplified using 100 ng, 10 ng, 1 ng or 100 pg of human genomic DNA as template. KAPATaq HotStart exhibits improved sensitivity, specificity, and yield when compared with competitive hotstart products. All reactions were performed using the manufacturer recommended protocols.

KAPATaq HotStart improves the specificity and sensitivity in PCR

A 270 bp amplicon was amplified from mycoplasma DNA with KAPATaq or KAPATaq HotStart. Sensitivity was tested using a 10x template dilution series starting with 1 ng of DNA. Higher sensitivity and less primer dimers are observed when using KAPATaq HotStart.

KAPATaq HotStart Kit Components

  • KAPATaq HotStart DNA Polymerase (5 U/µl in storage buffer)
  • 5x KAPATaq HotStart Buffer without MgCl2
  • 25 mM MgCl2
  • dNTP Mix (10mM each dNTP) KK1509 and KK1511 only
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