Robust DNA polymerase master mix

The Scripps Research Institute
Immunology Microbiology
Postdoctoral Researcher

Overall

Quality of Results

Ease-of-Optimization

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Company:

New England Biolabs

Product Name:

Q5® Hot Start High-Fidelity 2X Master Mix

Catalog Number:

M0494L

I work with numerous strain variants of a bacterium that require verification once in a while. Some genes across the strains exhibit variations in both DNA sequence and length. I amplify these genes using Q5 DNA polymerase and common oligos and validate them through gel electrophoresis, followed by DNA sequencing.

Experimental Design and Results Summary

Application

Amplification of DNA and cloning.

Starting Material

N/A

Tips

While working with the Q5 DNA polymerase master mix, always store it in a mini freezer. Avoid placing it on ice.

Results Summary

As shown in the figure, I performed colony PCR and separated the amplified DNA using agarose gel electrophoresis. As expected, I observed significant differences and the absence of DNA, confirming the authenticity of my Enterococcus strains. Also, I noticed that colony PCR fails with other master mixes; however, with the Q5 master mix, colony PCR never fails.

DOI or PMID #

N/A

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Summary

The Good

Highly reliable when compared to other cheap master mixes.

The Bad

Expensive.

The Bottom Line

The Q5 Master Mix (2X) from NEB provides a reliable solution for cloning purposes due to its outstanding quality. Strongly recommended.

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