Recommendations for Optimal Standard (End-Point)
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
4. Use a MasterAmp™ PCR Optimization Kit for optimal PCR using
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
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.
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.
d. RT activities have been reported for Taq and Tfl DNA
polymerases, but are, at best, too low to be useful.