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AllTaq Master Mix and PCR Core Kits

For ultrafast and versatile hot-start PCR in all applications
  • Simple, visual pipetting control and gel tracking dyes monitor successful procedure
  • Same 45-minute protocol for all targets including GC-rich and long targets up to 9 kb
  • Versatile chemistry enables duplex PCR and the inclusion of internal controls
  • 4x concentrated master mix and outstanding room temperature stability
  • Guard-protected, hot-start chemistry provides superior specificity and sensitivity

The AllTaq Master Mix Kit and the AllTaq PCR Core Kit provide a convenient format for highly sensitive and specific hot-start PCR using any DNA or cDNA template. Both kits are highly suited for all PCR applications and provide the following features: visual pipetting controls, gel loading and tracking dyes, an ultrafast cycling protocol, extreme room temperature stability during and after reaction setup and a 4x-concentrated master mix format, allowing a higher sample input volume.

Cat No./ID: 203123
AllTaq PCR Core Kit (250 U)
EUR 165,00
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50 µl AllTaq Polymerase (5 U/µl), 1.2 ml AllTaq PCR Buffer (5x), 55 µl dNTP Mix (10 mM each), 200 µl Template Tracer (25x), 50 µl Master Mix Tracer (125x), 2 ml Q-Solution (5x), 1.2 ml MgCl2 (25 mM), 1.9 ml RNase-Free Water
Cat No./ID: 203125
AllTaq PCR Core Kit (1000 U)
EUR 499,00
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200 µl AllTaq Polymerase (5 U/µl), 2 x 1.2 ml AllTaq PCR Buffer (5x), 200 µl dNTP Mix (10 mM each), 200 µl Template Tracer (25x), 2 x 50 µl Master Mix Tracer (125x), 2 ml Q-Solution (5x), 1.2 ml MgCl2 (25 mM), 4 x 1.9 ml RNase-Free Water
Cat No./ID: 203127
AllTaq PCR Core Kit (5000 U)
EUR 2.219,00
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5 x 200 µl AllTaq Polymerase (5 U/µl), 9 x 1.2 ml AllTaq PCR Buffer (5x), 5 x 200 µl dNTP Mix (10 mM each), 4 x 200 µl Template Tracer (25x), 9 x 50 µl Master Mix Tracer (125x), 5 x 2 ml Q-Solution (5x), 2 x 1.2 ml MgCl2 (25 mM), 16 x 1.9 ml RNase-Free Water
Cat No./ID: 203144
AllTaq Master Mix Kit (500)
EUR 356,00
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For 500 x 20 µl PCR amplifications: 2 x 1.25 ml AllTaq Master Mix (4x), 1 x 200 µl Template Tracer (25x), 2 x 50 µl Master Mix Tracer (125x), 5 x 1.9 ml RNase-Free Water
Cat No./ID: 203146
AllTaq Master Mix Kit (2500)
EUR 1.535,00
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For 2500 x 20 µl PCR amplifications: 10 x 1.25 ml AllTaq Master Mix (4x), 5 x 200 µl Template Tracer (25x), 10 x 50 µl Master Mix Tracer (125x), 20 x 1.9 ml RNase-Free Water

AllTaq Master Mix and PCR Core Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.


0
PCR buffer.
Ammonium and potassium cations in QIAGEN PCR Buffers increase specificity of primer annealing. K+ binds to the phosphate groups (P) on the DNA backbone, stabilizing the annealing of the primers to the template. NH4+, which exists both as the ammonium ion and as ammonia under thermal-cycling conditions, can interact with the hydrogen bonds between the bases (B), destabilizing the weak hydrogen bonds at mismatched bases. The combined effect of the two cations maintains the high ratio of specific-to-nonspecific primer-template binding over a wide temperature range.
1
No annealing temperature optimization.
PCR reactions were run using the AllTaq Master Mix Kit with different targets, varying template amount and annealing temperature (from 50°C to 60°C). Analysis was performed on the QIAxcel Advanced. All reactions resulted in successful and specific amplification of all targets.
2
Principle of AllTaq hot-start mechanism.
The DNA polymerase is kept in an inactive state by the antibody and the guard molecule until the initial heat activation step.
3
Single copy detection.
The gDNA template was diluted to theoretical one copy per reaction for amplification of CFTR (228 bp amplicon) using the AllTaq Master Mix Kit. A total of 68 parallel PCR reactions were run and then analyzed using the QIAxcel Advanced. 42 positive reactions were achieved experimentally, which is highly consistent with the 43 positive reactions predicted theoretically, using Poisson’s equation. Positive reactions are indicated with a "+", and negative reactions are indicated with a "–". This comparison demonstrates reliable and highly sensitive single copy detection with the AllTaq Master Mix Kit.
4
Amplification of long PCR fragments.
The AllTaq Master Mix Kit was used to amplify two DDX31 amplicons; 7644 bp and 8932 bp. Analysis was performed by agarose gel electrophoresis. Arrows indicate specific product.
5
Visual pipetting control.
The kit comes with optional pipetting controls. The Master Mix tracer is an inert orange dye that can be added to the Master Mix. The Template Tracer is an inert blue dye that can be added to the template. When adding the template to the Master Mix, the color turns green, providing a visual indication of correct pipetting. In addition, both dyes allow tracking during gel electrophoresis. The dyes run at about 50 bp (orange) and 4000 bp (blue) on a 1% agarose gel.
6
Integrated tracking dyes.
The AllTaq orange and blue tracer dyes serve as gel loading and fast-running (orange) and slow-running (blue) tracking dyes.
7
PCR procedure.
The AllTaq PCR Kit allows fast and easy PCR setup – just mix all components together in one tube and start your thermal-cycler program. The reaction mixture contains all of the reagents required for PCR. As a visual pipetting control, you can add the optional orange Master Mix Tracer and blue Template Tracer, verifying correct reaction setup, and allowing tracking during gel electrophoresis.
Performance
The innovative formulation of the AllTaq Master Mix and AllTaq PCR Buffer facilitates the amplification of specific PCR products and delivers successful results at the first attempt, using the same protocol for all targets. During the annealing step, the buffer allows a high ratio of specific-to-nonspecific primer binding (see figure PCR buffer). The verified buffer composition is adapted to ultra-fast cycling conditions and simultaneously provides stringent primer-annealing conditions over a wide range of annealing temperatures. The AllTaq PCR Buffer also ensures perfect duplex capabilities. Optimization of PCR by varying the annealing temperature (see figure No annealing temperature optimization) or the Mg2+ concentration is not required.

The versatile chemistry enables amplification of long targets up to 9 kb as well as duplex PCR, providing flexible workflow planning using the same PCR kit. Q-solution provided in the AllTaq PCR Core Kit facilitates amplification of difficult secondary structures, such as GC-rich templates. The guard-protected, hot-start mechanism (see figure Principle of AllTaq hot-start mechanism) prevents premature PCR leakage, ensuring premium specificity and sensitivity down to a single target molecule (see figure Single copy detection).
Principle
The AllTaq Master Mix Kit provides a convenient, ready-to-use master mix formulation. The AllTaq PCR Core Kit offers AllTaq DNA Polymerase, AllTaq PCR buffer, dNTPs, MgCl2 and Q-solution in separate tubes, in case optimization of the PCR protocol is desired (see figure Amplification of long PCR fragments). Both kits provide orange Master Mix Tracer and blue Template Tracer in separate vials for optional use.

At low temperatures, AllTaq DNA Polymerase is kept in an inactive state by an antibody and a novel guard molecule, which stabilize the complex (see figure Principle of AllTaq hot-start mechanism). This improves the stringency of the hot start and prevents any enzymatic activity at ambient temperature, allowing reaction setup without ice, facilitating automation. The enzyme is fully activated after the 2-minute incubation step at 95°C and starts amplifying with high specificity from the first cycle. The hot-start procedure eliminates extension from nonspecifically annealed primers and primer–dimers from the first cycle, ensuring highly specific and reproducible PCR.

The innovative AllTaq PCR Master Mix and AllTaq Buffer facilitate ultra-fast cycling conditions and simultaneously provide stringent primer-annealing conditions over a wide range of annealing temperatures. They also ensure perfect duplex capabilities. Optimization of PCR by varying the annealing temperature or the Mg2+ concentration is not required.

The blue and orange dyes in the PCR Template Tracer and in the PCR Master Mix Tracer, respectively, allow visual tracking of pipetted samples during the PCR setup, preventing errors. When the blue template is added to the orange PCR Master Mix, the color changes to green, confirming that sample was added (see figure 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 (see figure Integrated tracking dyes).
Procedure
The AllTaq Master Mix Kit provides a convenient, ready-to-use 4x concentrated master mix formulation, which allows for a high template input volume to further boost sensitivity. The AllTaq PCR Core Kit offers AllTaq DNA Polymerase, PCR buffer, dNTPs, MgCl2 and Q-solution in separate tubes, if optimization of the PCR protocol should be desired.

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 figure 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 figure Integrated tracking dyes).

The versatile buffer system allows ultrafast PCR, in approximately 45 minutes, using the same protocol for any standard PCR and duplex PCR. A cycling protocol is provided for long templates, between 1–9 kb.

The guard-protected, hot-start mechanism enables complete reaction setup at room temperature, eliminating the need for ice and allowing automated pipetting. After reaction setup, samples can be left at room temperature for more than 3 days before PCR cycling. Furthermore, the master mix is extremely stable, avoiding degradation during use and allowing storage at 2–8°C for up to 6 months.

The AllTaq PCR Core Kit contains Q-Solution that can be added optionally. Q-solution facilitates amplification of difficult targets by modifying the melting behavior of nucleic acids (e.g., templates with a high degree of secondary structure or are GC rich).

To further streamline PCR workflows, we recommend the QIAgility instrument for automated reaction setup and PCR analysis on the QIAxcel Advanced system.


Applications
The AllTaq Master Mix and AllTaq PCR Core Kits can be used for any PCR, for genotyping and genetic analysis or target detection from complex gDNA or cDNA templates, including reactions with multiple primer pairs or that include a wide range of amplicon sizes.

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