Choose the Right DNA Polymerase for PCR

Make sure you're using the optimal polymerase for your DNA amplifications. Get tips on choosing the right DNA Polymerase for your application.

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Fiona Stewart:

New England BioLabs has many polymerases designed for use in PCR. How do you decide which one you need? This will depend primarily on your application. The first decision you'll probably make is, do you need high fidelity amplification, or standard amplification? If you plan to use your PCR product in subsequent experiments such as protein expression or sequencing, you want to minimize the incorporation of errors, and you should choose a high-fidelity polymerase. If you just need to see if you have a PCR product or not, for example if you're confirming that your plasmid has an insert, then standard or colony PCR will be appropriate.

For high-fidelity amplification, we recommend starting with Q5 high-fidelity DNA polymerase, since it has the lowest available error rate, and has also been optimized for really robust performance across the ATGC spectrum. For standard PCR or colony PCR, we recommend starting with OneTaq DNA polymerase, which has also been optimized for great performance even with difficult templates such as GC rich.

Next, do you need to set up your reactions at room temperature? Is specificity a challenge? If so, a hot start version of the polymerase will be ideal.

Your next decision will be the polymerase format. For ease of use, we have master mixes that contain the enzyme, buffer, and dNTPs, and also quick load master mix formats that also contain loading dye so that you can load your PCR straight onto the gel. And of course, we also have stand alone polymerases.

For more specialized applications including amplification of next-generation sequencing libraries, very long PCR, or PCR direct from blood, we also have polymerase solutions. For guidance on which polymerase to select for your application, you can visit confidentPCR.com, or refer to the polymerase selection tools elsewhere on our website and in our marketing literature.

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