PCRBIO Ultra Polymerase & Mixes PCRBIO Ultra Polymerase & Mixes PCRBIO Ultra Polymerase & Mixes PCRBIO Ultra Polymerase & Mixes PCRBIO Ultra Polymerase & Mixes PCRBIO Ultra Polymerase & Mixes PCRBIO Ultra Polymerase & Mixes PCRBIO Ultra Polymerase & Mixes PCRBIO Ultra Polymerase & Mixes

PCRBIO Ultra Polymerase & Mixes

PCRBIO Ultra Polymerase is designed to amplify very challenging samples. Proprietary modifications increase progressivity, combined with advanced buffer chemistry and hot start technology, delivering excellent performance whether your template is high GC or AT, low abundance, or contains PCR inhibitors. Characteristics - Increase PCR success rate with challenging templates - Exhibits high tolerance to PCR inhibitors - Efficient specific amplification from complex templates including GC-rich sequences (up to 80%) - Antibody-mediated hot start allows for unrivaled detection from low copy number templates - Advanced buffer chemistry includes Mg and dNTPs - High yield under standard and fast PCR conditions - Three times higher accuracy than Taq - Conveniently available as a 2x ready-to-use mix with an optional red dye for direct gel loading.
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CAT NumberProductSizePrice
PB10.31-02PCRBIO Ultra Polymerase250 UnitsContact us
PB10.31-10PCRBIO Ultra Polymerase1000 UnitsContact us
PB10.32-01PCRBIO Ultra Mix80 x 50 μL ReactionsContact us
PB10.32-05PCRBIO Ultra Mix400 x 50 μL ReactionsContact us
PB10.33-01PCRBIO Ultra Mix Red80 x 50 μL ReactionsContact us
PB10.33-05PCRBIO Ultra Mix Red400 x 50 μL ReactionsContact us

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Additional Information

PCRBIO Ultra Polymerase is an extremely powerful enzyme, designed for efficient and reliable amplification of difficult and complex targets, even under challenging conditions such as the presence of inhibitors. The enzyme and buffer system have been developed to provide superior PCR performance and high success rates over a range of templates, including complex genomic DNA and high GC content targets (up to 80% GC).

PCRBIO Ultra Polymerase exhibits high tolerance to PCR inhibitors, making it an ideal choice for PCR from colony and crude samples. The PCR products generated have A-overhangs and can be cloned into TA cloning vectors. This enzyme comes with an optimized buffer that contains dNTPs and MgCl2.

Our antibody-intermediated hot start formulation prevents primer-dimer formation and non-specific products, allowing for specific and sensitive amplification from low copy number target sequences. For added convenience, PCRBIO Ultra Polymerase is also available as a 2x ready mix containing all reaction components except primers and template. PCRBIO Ultra Mix Red contains a preloaded red dye suitable for direct loading and tracking during agarose gel electrophoresis.

Applications

  • Challenging PCR - GC and AT-rich DNA
  • Low copy number template detection
  • PCR from crude samples
  • Colony PCR
  • Long-range PCR
  • Diverse PCR
  • TA cloning
  • Next-generation sequencing

Technical Specifications

PCRBIO Ultra Polymerase

Component 250 Units 1000 Units
PCRBIO Ultra Polymerase (5u/μl) 1 x 50μL 4 x 50μL
5x PCRBIO Ultra Buffer 2 x 1mL 8 x 1mL

PCRBIO Ultra Mix

Component 80 Reactions 400 Reactions
2x PCRBIO Ultra Mix 2 x 1mL 10 x 1mL

PCRBIO Ultra Mix Red

Component 80 Reactions 400 Reactions
2x PCRBIO Ultra Mix Red 2 x 1mL 10 x 1mL

Reaction Information

Reaction Volume Storage
50μL Upon arrival, the product should be stored between -30 to -20 °C. If properly stored, the kit will retain full activity until the expiry date indicated.

FAQ

The 5x PCRBIO Ultra Buffer supplied with PCRBIO Ultra Polymerase is specifically developed for this enzyme and we strongly recommend using them together. However, PCRBIO Ultra Polymerase should be compatible with any PCR buffer developed for use with wild-type Taq. If you use a custom buffer with PCRBIO Ultra Polymerase, keep in mind that reaction parameters such as annealing temperature and enzyme, template, dNTP, and MgCl2 concentration may need optimization.

Yes. If you are working from bacterial colonies, use a sterile needle to take a colony and suspend it into the 50µl PCR reaction. If working from liquid culture, add 5µl of overnight culture into the final mixture. Follow the general protocol and increase the initial denaturation time to 10 minutes at 95°C.

Yes. Use 2 µL of blood sample for a 50 µL PCR reaction and follow the general protocol. Please note that components in the blood may inhibit the PCR. Perform a dilution series on the sample to determine the optimal template concentration for PCR amplification.

If smearing is a problem, the best approach is to ensure that it is not an artefact of running an agarose gel under suboptimal conditions. Suboptimal conditions include high voltage or not allowing enough time for the gel to set1.

You may also need to troubleshoot the PCR reaction and consider the following suggestions2.

  • Primers should be designed to prevent primer-primer interactions and improve specificity.
  • Increase the annealing temperature or perform PCR with an annealing temperature gradient to find the optimal annealing temperature.
  • Reduce the amount of template in the reaction. For high-quality DNA, use 1–100 ng of genomic DNA or ≤5 ng of plasmid/lambda DNA per 50 µL reaction.
  • Reduce the number of cycles.
  • Reduce the amount of enzyme per reaction.
  • Decrease the primer concentration, but do not go below 100 nM of each primer.
  • Add DMSO into the reaction with a final concentration of 5%–10%.

1  Koontz, L. Agarose gel electrophoresis. Methods in enzymology: DNA. 1st edn, Vol. 529 35-45 (2013).

2  Lorenz, T. C. Polymerase chain reaction: basic protocol and troubleshooting and optimization strategies. J Vis Exp, e3998, doi:10.3791/3998 (2012).

You may want to consider the suggestions below and also consult the literature1.

  • Optimize the annealing temperature for PCR with a gradient of annealing temperatures.
  • Increase the amount of template in the reaction.
  • Increase the number of cycles.
  • Increase the amount of enzyme per reaction.
  • Increase the primer concentration, but do not exceed 1 µM of each primer.
  • Try a fresh dNTP solution.
  • Optimize MgCl2 concentration.

1  Lorenz, T. C. Polymerase chain reaction: basic protocol and troubleshooting and optimization strategies. J Vis Exp, e3998, doi:10.3791/3998 (2012).

This non-inhibitory dye, added to allow for direct gel loading, migrates at a similar rate to 200-300 bp DNA fragments on a 1% agarose gel and at 50-100 bp on a 2% agarose gel. You may notice a shift in this apparent molecular weight when running gels with varying agarose content.

This enzyme has an error rate of approximately 1 error per 5.0 x 105 nucleotides incorporated.

For products under 5kb, we recommend using 15 seconds per kb when amplifying from eukaryotic DNA. For longer products, we recommend extension times of 40 to 60 seconds per kb.
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