PCR Master Mixes

PCR Master Mixes

Our ready to use Master Mixes are great alternative options to your regular individual PCR reagents. Master mixes are reagents that include all the components needed for a PCR. By only the extra step of adding your template and primer pairs you are able to run a PCR quickly and successfully.

  • Minimize possible contamination of the stock reagents due to fewer handling steps, enhancing the reproducibility of your experiments. 
  • Reduce your experiments time

Each of our Master Mixes has its own benefits. All the master mixes specified below are available with different buffer options, giving wider options to finally chose the master mix that matches your PCR conditions and application. 

 

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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 and are available with a final MgCl2 concentration of 1.5 mM or 2.0 mM.

  • Reduce the extra  step of adding loading buffers: The color red of the master mix provides a perfect direct loading of your PCR products after amplification. It enables the visualization of the product when pipetting and gel loading.

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 a stabiliser, which do not affect  PCR reaction.

If desired for your protocol experiments, the red dye can be removed afterwards by using PureIT ExoZAP PCR CleanUp or by spinning the column purification. Moreover, with your clean PCR product, your sample can be used for sequencing.

 Taq DNA Polymerase Master Mix RED

Features

  • Premixed solution
  • Direct loading on agarose and SDS DNA gels
  • Easy visualization of pipetting and loading gels
  • Minimal optimization
  • 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

Our Taq DNA Polymerase Master Mix RED showed great performance and results when targeting targets different length genes: PAH (203 bp), Q8 (727 bp), CFTR (613 bp) and BAIP (788 bp).

Direct gel loading

 

Applications: 

Genotyping using Q-Extract DNA Extraction PCR Kits

The AppNote shows the study performed with the Q-Extract DNA Extraction PCR Kits in combination with either one of the two available master mix: Taq DNA Polymerase 2x Master Mix RED or TEMPase Hot Start DNA Polymerase 2x Master Mix BLUE. 

The extraction and amplification of genomic DNA was performed from five different mouse tissues (kidney, muscle,
lung, liver and tail), chicken muscle tissue and human saliva. 

  • The results show that cam be used for many different mammalian samples and also saliva.
  • A wide range of sample amounts was analysed 
  • Genotyping results were achieved in 70 to 210 minutes 

Read to AppNote for more information about the use of the master mix. 

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  nice technique that enables the quantification of DNA and the analysis of gene expression levels. 
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. Additionally, our 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. As a result, our master mix provides you reliable and robust results time after time.

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

  • RealQ Plus Master Mix Green: DNA-binding fluorescent with a green dye for detection.
  • RealQ Plus Master Mix for Probe: it uses a detection based on probe real-time PCR.

 

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

To select the correct ROX™ reference dye levels for your specific Real-Time PCR applications, check the “Real-Time instrument guide”. An advantage of our two variants of master mixes is that both can be chosen when many genes need to be analysed within a short amount of time. 

Do you want to know the specific ROX™ reference dye levels (without rox, with low rox, and with high rox) that matches better your experiment? 

Contact us 

 

Stability studies with the qPCR master mix

Experiment: 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.

Results: High stability and complete inactivation state of the TEMPase is seen before hot start. This allows researchers to run their plates at room temperature, avoid changes on temperature, and work on a more convenient time instead of directly after setting up the reaction.

Room temperature stability 48hRoom temperature stability 48h

Room temperature stability 72hRoom temperature stability 72h

AQ97 High Fidelity DNA Polymerase Master Mix

AQ97 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. .60 x Taq fidelity
•    3’ to 5’ proofreading exonuclease activity
•    High elongation rate: 10sec/kb
•    Long-range capability: 11kb gDNA
•    Good amplification on DNA template with low to high GC content

 

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DITRIBUTION OF SUBSTITUTION ERRORS

PCR was performed using Taq DNA Polymerase, AQ97 High Fidelity DNA Polymerase, high fidelity DNA Polymerase Q and high fidelity DNA Polymerase P.

The PCR was followed by NGS sequencing of the PCR products. The number of substitutions at each PCR target position was calculated and plotted in diagram A. Substitutions include misincorporated nucleotides and deletions at each position. Non-polymerase errors are subtracted from the total number of errors to revel true polymerase errors. Non-polymerase errors include mutations caused by thermocycling-induced DNA damage, pre-NGS sample preparation and sequencing errors. In these diagrams the average number of substitutions for Taq DNA Polymerase (Taq average) and for AQ97 High Fidelity DNA Polymerase (AQ97 average) is also plotted. Diagram B magnifies the area near the detection limit, displaying more information about the number of substitutions for AQ97 High Fidelity DNA Polymerase, high fidelity DNA Polymerase Q and high fidelity DNA polymerase P.

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

Subcategory

Unit

Code
Description

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