PCR Master Mixes

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.

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).

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.