Best PCR Master Mix Available

December 02, 2015

University of North Carolina at Chapel Hill
Research
Laboratory Technician

Overall

Quality of Results

Ease-of-Optimization

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Company:

Bioline

Product Name:

ImmoMix Red 2X

Catalog Number:

BIO-25022

Our lab conducts a great deal of translational research with transgenic mice, and we run a variety of PCR's to genotype mice. We rely on a singular PCR Master Mix when genotyping all of our transgenic alleles because this particular polymerase is of extremely high efficiency and specificity.

Experimental Design and Results Summary

Application

PCR Reactions

Starting Material

Transgenic Mouse DNA

Protocol Overview

Preparing PCR Reactions is extremely fast with this product. Because this is a Master Mix, we only need to add primers, water, and potentially 5% DMSO for each reaction. In addition, it comes with loading dye/buffer included, so you don't need to add loading buffer to each sample before running out the reactions on an agarose gel. Heat activation of the polymerase only takes a few minutes on the thermal cycler.

Tips

We use a final reaction volume of 10 uL instead of the recommended 20 uL, with no difference whatsoever.

Results Summary

ImmoMix gives very clean bands of DNA once run out on a gel, with very few exceptions. Several of our PCR's will only work when using this PCR Master Mix, likely because of the quality of the Polymerase. Furthermore, it's easy to see the dye front on the gel even when run out for hours.

Additional Notes

Not having to add loading buffer to each tube post-PCR will save you time and effort.

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Summary

The Good

Ease of use, quality of polymerase, reliability of product following multiple freeze-thaws

The Bad

Expensive....also the salt concentration is rather high, and can't be altered.

The Bottom Line

This is our lab's go-to PCR Master Mix. While it is expensive, it consistently outperforms Polymerases and PCR Master Mixes from other companies and does so with an increased ease of use. We have been using this Master Mix for almost 10 years now without any need to change. Highly recommended.

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