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UCP HiFidelity PCR Kit

UltraClean Production master mix for high-fidelity hot-start PCR and microbiome applications 

  • Eliminate residual DNA and background noise with ultra clean, DNA-depleted reagents
  • Acquire reliable, robust results without inhibitors over wide GC-content range 
  • Exceptionally fast PCR with fidelity 70 times higher than Taq
  • Superior specificity and sensitivity: antibody-mediated hot start of polymerase and exonuclease
  • Visual pipetting control and gel tracking dyes monitor successful procedure
  • Automation ready: stable, room-temperature reaction setup

The UCP HiFidelity PCR Kit provides a convenient format for highly sensitive and specific hot start PCR that need to have low PCR-error rates using any DNA or cDNA template.

 

Dedicated processes enable ultra clean production of this master mix. These processes deplete potential background DNA to provide the cleanest sample possible. Combined with superior inhibitor resistance and minimized GC-content bias, this kit is an ideal choice for microbiome applications such as 16S or 18S amplification. This is complemented with a 70 times higher fidelity compared to Taq polymerase, thus enabling error-free amplification, as needed during cloning, amplicon-sequencing or analysis of genome-editing experiments. Additional features include visual pipetting controls, a fast cycling protocol, room-temperature stability during and after reaction setup and the ability to amplify long amplicons.

Want to try this solution for the first time? Request a quote for a trial kit.

 

 

Cat No./ID: 202742
UCP HiFidelity PCR Kit (100)
EUR 157,00
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For 100 x 25 µl PCR reactions: 1 x 1.3 ml UCP HiFidelity MasterMix (2x); 1 x 200 μl UCP Template Tracer, 25x; 1 x 50 μl UCP Master MixTracer, 125x; 1 x 1.9 ml UCP PCR Water
Cat No./ID: 202744
UCP HiFidelity PCR Kit (500)
EUR 655,00
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For 500 x 25 µl PCR reactions: 5 x 1.3 ml UCP HiFidelity MasterMix (2x); 2 x 200 μl UCP Template Tracer, 25x; 1 x 50 μl UCP Master MixTracer, 125x; 4 x 1.9 ml UCP PCR Water

This product is licensed from Bio-Rad Laboratories, Inc., under U.S. Pat. Nos. 6,627,424, 7,541,170, 7,560,260 and corresponding patents in other countries for use solely in DNA sequencing, DNA micro-array, and conventional PCR applications, including pre-amplification steps that are required for such applications, in the life science research and in-vitro diagnostics fields but not real-time PCR or digital PCR.


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Amplify high and low GC-content templates with no protocol or buffer optimization.
Templates with high, medium and low GC content [85% (MCART), 65% (PROM1) and 25% (PROM410)] were successfully amplified using the same standard protocol and buffer conditions. The UCP HiFidelity PCR Kit prevents amplification bias due to GC content with no optimization required.
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Easy visualization with orange and blue tracking dyes.
The dyes run at approximately 50 bp (orange) and 4000 bp (blue) on a 1% agarose gel.
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Exceptional inhibitor robustness enabling successful amplification of various targets directly from whole blood.
Seven targets varying in size and composition were amplified directly from 1 µl whole blood with no reaction optimization.
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PCR procedure
The orange Master Mix Tracer is optionally added to master mix or PCR buffer. When template dyed with the blue Template Tracer is added to the master mix, the color of the solution changes from orange to green, providing a visual indication of correct pipetting and reaction setup. The Tracer dyes allow direct loading on agarose gels after PCR, and efficient tracking of the subsequent electrophoresis. The dyes run at approximately 50 bp (orange) and 4000 bp (blue) on a 1% agarose gel.
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Premium fidelity compared to competitors for the most reliable gene editing and sequencing.
The UCP HiFidelity PCR Kit outperformed the competition with fidelity, 70x greater than Taq, in the rigorous Lacl Fidelity test.
5
Sensitive bacterial and fungal amplification with no background ideal for low biomass microbial samples.
As little as 0.01 pg E. coli and 0.05 pg Candida were successfully detected and NTC showed no background.
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Superior inhibitor resistance compared to competitors.
A 2 kb fragment of human HRPT1 was amplified using the UCP HiFidelity PCR Kit and competitor master mixes, following the recommended protocols and using a 25 µl final reaction volume. Reactions were amplified using 10 ng human gDNA. Parallel reactions were run, with no inhibitor (N.I.), with 25 ng humic acid (H.A.), 25 µM Heparin (H), 25 ng bile salt (B.S.) and with no template control (NTC). Analysis was performed using the QIAxcel. Only the UCP HiFidelity PCR Kit amplified the specific product in the presence of all inhibitors.
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UCP HiFidelity PCR Kit
For 100 x 25 µl PCR reactions: 1 x 1.3 ml UCP HiFidelity MasterMix (2x); 1 x 200 μl UCP Template Tracer, 25x; 1 x 50 μl UCP Master MixTracer, 125x; 1 x 1.9 ml UCP PCR Water
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Versatile detection of various templates with amplification over a wide range of annealing temperatures.
18S and 16S fragments were successfully amplified with annealing temperatures ranging from 55-72°C, using constant denaturation (98°C) and extension temperatures (72°C) over 35 cycles. The UCP HiFidelity PCR Kit demonstrates the tremendous flexibility of its reaction buffer.
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Visual pipetting control
When the blue template is added to the orange PCR Master Mix, the color changes to green, confirming that sample was added.
Performance

Ultra-Clean Production processes combined with nucleic acid depletion ensure potential contamination with residual DNA from expression host or other sources is reduced to a minimum. All components are tested for bacterial and fungal DNA, using generic PCR Assays for 16S and 18S rRNA coding sequences. This facilitates accurate results in microbial testing, quality control procedures or microbiome / metagenome analysis and sequencing (See: "Sensitive bacterial and fungal amplification"). Low biomass sample testing, in particular, benefits from ultra clean reagents and with improved inhibitor resistance and minimal GC-content bias ensures accurate results (See: "Superior inhibitor resistance..." and "Amplify high and low GC…").

 

The enzyme’s proofreading feature provides a significantly improved fidelity, 70 times higher than the standard Taq polymerases it was compared against (See: "Premium fidelity").

 

Both the polymerase activity and the exonuclease activity are stringently controlled by specific antibodies, thus preventing premature activity or damage and nonspecific annealing of template and primers during reaction setup. The kit creates blunt end products and is perfectly suited for procedures requiring high-fidelity PCR such as cloning, site directed mutagenesis, overlap extension PCR or analysis of gene editing experiments.

 

The capability of low degree multiplexing and amplification of long targets up to 10 kb also makes it a perfect choice for amplicon sequencing analysis. Room temperature set-up, integrated visual pipetting and gel loading dyes further improve efficiency in genotyping and genetic testing workflows.

 

Principle

The UCP HiFidelity PCR Kit provides a convenient ready-to-use master mix formulation and UCP PCR water. It also contains orange UCP Master Mix Tracer and blue UCP Template Tracer in separate vials for optional use (see: "UCP HiFidelity PCR Kit"). The enzyme’s proofreading feature provides a significantly improved fidelity that is 70 times higher when compared to standard Taq polymerases. Both the polymerase activity and the exonuclease activity are stringently controlled by specific antibodies, thus preventing premature activity or damage and nonspecific annealing of template and primers during reaction setup. At low temperatures, the enzyme’s polymerase and exonuclease activities are kept in an inactive state, thus preventing any enzymatic activity at ambient temperatures and until heat activation at 98°C.

 

The blue and orange dyes in the UCP Template Tracer and in the UCP Master Mix Tracer, respectively, allow visual tracking of pipetted samples during the PCR setup, to prevent errors. When the blue template is added to the orange PCR Master Mix, the color changes to green, confirming that sample was added (See: "Visual pipetting control". The use of these tracers is optional.

 

Reactions can be directly loaded onto agarose gels after cycling. Each tracer dye allows monitoring of the loading process and efficient tracking of the subsequent electrophoresis. The blunt ended PCR-product is highly suited for various cloning or gene editing applications.

 

Procedure

The UCP Multiplex Kit provides a convenient ready-to-use 2x concentrated master mix formulation.

 

The orange Master Mix Tracer is optionally added to master mix or PCR buffer. When template dyed with the blue Template Tracer is added to the master mix, the color of the solution changes from orange to green, providing a visual indication of correct pipetting and reaction setup (See: “PCR Procedure”). The Tracer dyes allow direct loading on agarose gels after PCR, and efficient tracking of the subsequent electrophoresis. The dyes run at approximately 50 bp (orange) and 4000 bp (blue) on a 1% agarose gel (See: “Easy visualization”).

 

The versatile buffer system allows ultrafast PCR in less than 1 hour without the need for optimization or changing buffer components (See: "Versatile detection…"). The kit additionally offers a protocol for amplification of long templates or low degree of multiplex PCR.

 

To further streamline PCR workflows, we recommend the QIAgility instrument for automated reaction set up, and PCR analysis on the QIAxcel system. For microbiome analysis, QIAGEN offers a range of dedicated products including DNeasy Power Soil Pro Kits for sample extraction, QIAseq 16S panels for next generation sequencing or CLC Microbial Genomics Pro Suite software for bioinformatic analysis.

 

Applications
The UCP HiFidelity PCR Kit is ideally suited for procedures requiring a high purity and accuracy, like microbial testing, microbiome/metagenome analysis and sequencing, quality control, genotyping and gene editing. The kit creates blunt-end products and is perfectly suited for procedures requiring high-fidelity PCR, such as cloning or site directed mutagenesis. The capability of low degree multiplexing and amplification of long targets up to 10 kb also makes it a perfect choice for amplicon sequencing analysis. The high inhibitor robustness even allows direct PCR from blood (See: "Exceptional inhibitor robustness...").

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