PCR Product Selection Guide

Recommendations for Optimal Standard (End-Point) PCR

1. Use the FailSafe™ PCR System to amplify any template up to 20 kb, whatever the sequence.

Whatever the template, whatever its sequence, the FailSafe™ PCR System is almost always your best choice for dependable, and consistent amplification of any template up to 20 kb. It is also highly accurate. The FailSafe™ PCR Enzyme Mix has a nucleotide incorporation fidelity at least 3-4 times better than Taq and hot-start PCR is not necessary with FailSafe protocols. Even with room-temperature set-up, FailSafe gives high specificity and excellent amplification without hot-start. Since we are cautious scientists, we were initially reluctant to use the name "FailSafe," but as we observed one difficult template after another being successfully amplified, that reluctance disappeared. It also helped our confidence to have so many people come to our booth at scientific conferences to tell us how the FailSafe System saved some aspect of their project. Try it next time a template cannot be amplified. Maybe you still have such a template in your freezer. The FailSafe PCR System amplifies any template, including those with high-GC sequences or other difficult structures that cause DNA polymerase stops or pausing. It also works great for multiplex PCR. At Epicentre, we use it for all templates <20 kb.

2. Use TAQXpedite PCR System with your FAST or conventional Thermal Cycler for reduced reactions times and increased productivity.

The TAQXpedite system offers a high fidelity enzyme with a Universal and Difficult/High GC MasterMix including our patented PCR Enhancer with Betaine. The enzyme is capable of 500bp in a little as 16 minutes with a standard thermal cycler.

3. Use the MasterAmp™ Extra-Long PCR System for templates greater than 20 kb.

This kit is specifically optimized for high-fidelity amplification of long templates - from 20 kb to 40 kb or more. It uses an enzyme mix and 9 MasterAmp™ 2X PCR PreMixes (different than those for the FailSafe System) that are specially formulated for long templates. Being cautious scientists, we did not feel comfortable to call this kit "FailSafe" with respect to every template from 20 to 40 kb or more. However, we do feel comfortable saying that there is no better product for amplifying long sequences.

4. Use a MasterAmp™ PCR Optimization Kit for optimal PCR using other enzymes.

While you will obtain the best PCR results by using the FailSafe PCR System for templates up to 20 kb and the MasterAmp Extra-Long PCR Kit for templates >20 kb, we also offer other thermostable DNA polymerases and 2 different MasterAmp™ PCR Optimization Kits. Both kits contain meticulously optimized enzyme-specific 2X PCR PreMixes and include our patented PCR Enhancer with Betaine so that you will obtain the best possible PCR results.

Use the MasterAmp™ PCR Optimization Kits WITH Ammonium Sulfate - For Tfl, Tth or Pfu-type DNA polymerases.

Use the MasterAmp™ PCR Optimization Kits WITHOUT Ammonium Sulfate - For Taq or AmpliTherm™ DNA Polymerase.

  5´→3´ structure-dependent exonuclease 3´→5´ proofreading exonuclease Terminal transferase activitya Reverse transcriptase activityc,d Amplify <3 kb DNA Amplify 3-5 kb DNA Amplify 5-15 kb DNA Amplify 20 kb DNA Amplify 40 kb DNA
FailSafe PCR System + + +b - + + + + -
MasterAmp Extra-Long PCR Kit + + +b - + + + + +
MasterAmp Taq DNA Polymerase + - + - + + + - -
MasterAmp Tfl DNA Polymerase + - + - + + + - -
MasterAmp Tth DNA Polymerase + - + + + + - - -
MasterAmp AmpliTherm DNA Polymerase - - + - + + - - -
TAQXpedite PCR System (FAST End-Point) + + +b - + + + + -

a. 3´-uncoded nucleotide addition (e.g., 3´-terminal A overhang).
b. PCR products made using these enzymes can be efficiently cloned in both blunt-end and 5´-T overhang vectors.
c. Mn2+-dependent.
d. RT activities have been reported for Taq and Tfl DNA polymerases, but are, at best, too low to be useful.