PCR Master Mix With Loading Dye

September 22, 2015

Duke University Medical Center
Associate Professor

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review

Company:

Genesee Scientific

Product Name:

Apex™ Taq RED Master Mix, 2.0X

Catalog Number:

42-137

I have used Taq Master Mix from Apex (sold by Genesee Scientific) for several years and have always had excellent results. It essentially always works and requires minimal optimization. I recently purchased Apex's RED Master Mix 2.0X, which is ready to load on an agarose gel directly from the PCR reaction. I performed a side-by-side comparison with the conventional Apex Master Mix and saw no difference in performance between the two.

Experimental Design and Results Summary

Application

End point PCR

Starting Material

DNA isolated from cells, plasma, and FFPE sections

Protocol Overview

Reactions included 30-50 ng DNA template. PCR conditions consisted of an initial denaturation at 94°C for 2 min, followed by 35 cycles of 15 sec at 94°, 30 sec at 56°, 45 sec at 72°, and final elongation for 5 min at 72°. The total PCR reaction volume was 25 µl.

Tips

Mix the RED Master Mix with primers and dispense into PCR tubes on ice. Add template and water to get a final volume of 25 µl.

Results Summary

After the PCR reaction, I loaded 10 µl of each reaction directly from the PCR tube onto a TAE agarose gel containing 1.7% agarose. The figure shows clean bands at the expected size. I purified the amplicons from this gel and had them sequenced. This verified that the expected amplicon was indeed produced in the reaction.

Additional Notes

If you are careful to design good primers, and your template is not degraded, the Apex RED Master Mix should work. I use the NCBI primer design application available free of charge on the NCBI site. If needed, a gradient of annealing temperatures can be run to optimize the reaction.

Related Categories

Image Gallery

Summary

The Good

Excellent performance and very economical.

The Bad

None

The Bottom Line

Apex Master Mixes have always worked extremely well for me. The inclusion of loading dye in the Master Mix simplifies the analysis of PCR reactions without affecting the quality of the product.

Join the discussion