PCR Master Mixes

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Our ready to use Master Mixes are great alternative options to your regular individual PCR reagents. By only adding your template and primer pairs you are able to run a PCR quickly and successfully. Due to the fewer handling steps contamination of the stock reagents is minimised and the reproducibility of your experiments will be enhanced. Next to this, it is simply less time consuming! 

Each of our Master Mixes has its own benefits. 
Scroll down to see our detailed overview. 

  • Taq DNA Polymerase Master Mix RED
  • RealQ Plus PCR Master Mix
  • AQ90 High Fidelity DNA Polymerase Master Mix
  • Amplidiag® Multiplex PCR Master Mix
  • Other Master Mixes

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Taq DNA Polymerase Master Mix RED

Ready-to-use premixed solutions are perfect for your standard PCRs. Taq polymerases give steady and reliable results. All Master Mixes are 2x reaction mixtures available with a final MgCl2 concentration of 1.5 mM or 2.0 mM.

Perfect for direct loading of your PCR products after amplification. No need for additional loading buffers. This Master Mix consists of a Taq DNA Polymerase, ammonium (NH4+) buffer system, dNTPs and magnesium chloride (MgCl2). 
Next to this, it contains a red dye and stabiliser which both do not affect the PCR reaction. If desired, the red dye can be removed afterwards by using PureIT ExoZAP PCR CleanUp or spin column purification. 
The amplified PCR products generated by Taq DNA Polymerase Master Mix RED can also directly be used for sequencing after treatment with the same methods used to remove the red dye. 

Features
•    Premixed solution
•    Direct loading on agarose and SDS DNA gels
•    Easy visualization of pipetting and loading gels
•    Minimal optimisation
•    Dye front runs at 300 – 1000 bp on a 0.5 – 1.5 % agarose gel
•    No need for a separate loading buffer
•    Higher reproducibility due to less pipetting steps
•    High product yield
•    dUTP incorporation possible
•    Processes up to 5 kb
•    Leaves a 3’dA overhang

DNA Polymerase Master Mix Red

Direct Gel Loading

PCR products can be loaded onto agarose and SDS DNA gels directly after amplification. The red dye visualizes the product for both pipetting and gel loading.

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Comparison

The Taq DNA Polymerase Master Mix RED was compared to other polymerases of well-known companies. Four DNA targets of different lengths were investigated; PAH (203 bp), Q8 (727 bp), CFTR (613 bp) and BAIP (788 bp). Taq DNA Polymerase Master Mix RED showed an equally well of better results in this study. Every amplification has been performed according to the suppliers protocol. 

RealQ Plus PCR Master Mix

RealQ Plus PCR Master Mixes are ready-to-use 2x master mixes used for different Real-Time PCR applications. Real-Time PCR is a very nice technique to analyse gene expression levels and quantification of DNA. 
RealQ Plus master mixes consist of TEMPase Hot Start DNA Polymerase, dNTPs, MgCl2 and a special buffer system necessary to carry out high quality DNA amplification. The RealQ Plus PCR Master Mixes have a high stability and efficiency when working with cDNA and gDNA. 
To ensure you that RealQ Plus PCR Master Mix gives the same robust results time after time, we can proudly say that the Master Mix is produced under ISO 9001:2015 conditions. 

Two variants are available in combination with three different ROX™ reference dye levels. 

RealQ Plus Master Mix Green; DNA-binding fluorescent dye detection
RealQ Plus Master Mix for Probe; probe-base real-time PCR based detection

To select the correct ROX™ reference dye levels for your specific Real-Time PCR applications, check the “Real-Time instrument guide”.
Both Master Mixes can be chosen when many genes need to be analysed within a short amount of time. 

Features
•    High specificity, reproducibility, efficiency and stability
•    Reaction set-up at room temperature
•    All-in-one premixed 2x solution

Amplification plot shows a 4-fold dilution series for the Pthr gene (75 bp); 80 ng to 80 pg of gDNA. RealQ Plus 2x Master Mix for Probe, High ROX™ was used. Samples are in triplicates. Results show a high efficacy and high precision which is close to 100%. 

This amplification plot shows high reproducibility capacity of the RealQ Plus 2x Master Mix Green, High ROX and 20 ng gDNA. 
Only 0.084 standard deviation after 80 replication

Stability

 

 

 

 

 

 

Two plates were prepared for a qPCR reaction, but were incubated afterwards in the dark at room temperature for 48 and 72 hr before the actual run. High stability and complete inactivation state of the TEMPase is seen before hot start. This allows scientists to run their plates at a later and/or more convenient time instead of directly after setting up the reaction.

 

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AQ90 High Fidelity DNA Polymerase Master Mix

AQ90 High Fidelity DNA Polymerase Master Mix is a 2x Master Mix used in applications in which high fidelity is important like in cloning, next generation sequencing (NGS) applications and/or mutagenesis. It has the ability to amplify difficult DNA targets (with low or high GC content) as well as long DNA targets. 

Features
•    2x all in one master mix
•    Quick reaction set up
•    High fidelity
•    3’ to 5’ proofreading exonuclease activity
•    Extra low error rate
•    Processes up to 8.5 kb gDNA and ≤ 12 kb for λDNA
•    Good amplification on DNA template with low to high GC content

Both AQ90 High Fidelity DNA Polymerase as well as AQ90 High Fidelity 2x Master Mix result in good results on various DNA targets (from low to high GC content). Above 9 different targets were tested, varying in length (200 – 700 bp) and GC content (29 – 78 %). Amplification of targets containing 71 – 78 % GC content was only working properly in combination with Betaine Enhancer Solution (2M) in the PCR mixture. 

 

Large and difficult amplicons can successfully be amplified by using the AQ90 High Fidelity DNA Polymerase 2x Master Mix as depicted here on the left. Four different human gDNA targets were used varying from 2 kb to 8.5 kb were used in this test.

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Amplidiag® Multiplex PCR Master mix


Amplidiag® Multiplex PCR Master mix is a 2x concentrated master mix with high performance and ready to use. The Master Mix is successfully tested for high specificity and sensitivity multiplex as well as singleplex real-time PCRs and PCR assays. Residual DNA levels are almost neglectable, which is an advantage for complicated applications. The Master Mix contains a Taq polymerase with hot start technology; low activation before hot start activation which increases the PCR specificity. 

Features

  • Good amplification in multiplex with high sensitivity of low frequent targets
  • Good amplification and extremely high sensitivity in inhibitory sample materials
  • Ultra-low residual DNA levels, ideal for bacterial DNA detection in multiplex assays

No Cq lag in multiplex assays

100 copies of Shiga toxin type 2 (stx2) ampliqons were amplified as well as 1000 copies Internal Amplification Control (IAC) ampliqons and multiple amounts of Shiga toxin type 1 (stx1) ampliqon copies (106, 107 and 108). As a comparison, the same ampliqons and amount were also run separately. When combining all results, as shown above, no differences in amplification is seen. Meaning no Cq lag for the different targets comparing multiplex to single plex runs. 
Labels used: FAM for stx1, HEX for stx2 and Cy5 for IAC.

High performance even after 24 h in RT

Below results are shown of a comparison experiment; Amplidiag Multiplex PCR Master mix versus a non-hot start Taq polymerase. Before running the PCRs, the complete reaction mixtures were pre-incubated for either 2 h or 24 h at room temperature. 

Amplification results (in duplicates) after pre-incubation periods. The end point intensities and Cq values show that the Amplidiag Mulitplex PCR Master mix (AD) is not affected compared to the non-hot start enzyme (T) which show very poor amplification

Results of a gel run (in duplicates) of the same reactions as described previously after amplification. 
On the left: results of the Amplidiag Mulitplex PCR Master mix (AD) which do not show loss of amplification of 100bp target. 
On the right: results of the non-hot start enzyme (T) which again show a failed amplification. 

Other Master Mixes:

  • Taq Polymerase Master Mix
    • Taq DNA Polymerase Master Mix 
    • Taq OptiMix CLEAR
  • TEMPase Hot Start DNA Polymerase Master Mix
    • TEMPase Hot Start Master Mix
    • TEMPase Hot Start Master Mix Blue
    • Multiplex TEMPase Master Mix
  • GC TEMPase Master Mix
    • GC TEMPase Master Mix l
    • GC TEMPase Master Mix ll
    • GC-rich DNA Target Kit

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