There are many kits and thermostable DNA polymerases available for PCR. What should you consider in order to choose which one is best for your application?

Generally speaking, a PCR system should possess at least three basic capabilities:

1. The product must be sensitive enough to amplify your DNA
2. The amplification must be accurate
3. The product must be able to amplify a DNA fragment as long as you desire

However, for a long time, a PCR system that was dependable, accurate and consistent in amplifying a variety of templates was considered impossible. For some templates, amplification either could not be achieved or it was accomplished only after spending a lot of time and work for optimisation of reaction conditions. Some DNA templates were too difficult to amplify because they were highly rich in GC content. In other cases, mis-priming or secondary structure hampered specificity, as well as sensitivity. Most enzymes, including Taq DNA polymerase, could not amplify long templates due to DNA polymerase pausing, low processivity, and inadequate fidelity. Furthermore, when amplification was achieved, PCR products often did not have the correct sequence, leading to lost time and effort. For example, Taq DNA polymerase does not synthesise DNA without errors, in part due to its lack of 3'-to-5' proofreading activity. These and other problems encountered with PCR have been resolved by the introduction of the FailSafe™ PCR System and the MasterAmp™ Extra-Long PCR System.

Two Proprietary Systems Enable Consistent, Dependable, Accurate PCR Results

The FailSafe™ PCR System produces consistent, dependable high fidelity PCR results for templates up to about 20kb, whatever its sequence or structure. For longer templates, from 20kb to about 40kb or more, the MasterAmp™ Extra-Long PCR Kit provides dependable and accurate amplification.

Unique Enzyme Blends Improve Fidelity & Sensitivity of PCR

The FailSafe™ and MasterAmp™ Extra-Long PCR Systems use proprietary blends of thermostable DNA polymerases that are capable of amplifying even the most difficult templates with extremely high sensitivity and high fidelity. Both of these unique enzyme blends synthesise DNA with fidelities >3 times higher than Taq DNA polymerase.

New Patented PCR Enhancer Technology Significantly Improves PCR Performance

Besides the unique enzyme blends, both the FailSafe™ and MasterAmp™ Extra-Long PCR Systems also incorporate Epicentre®'s PCR Enhancement Technology into the provided sets of FailSafe™ PCR PreMixes and MasterAmp™ Extra-Long PCR PreMixes, respectively. These PCR PreMixes include everything you need for successful PCR or Extra-Long PCR with all of your templates and primer pairs, including dNTPs, buffer, and varying amounts of MgCl2 and FailSafe or MasterAmp™ PCR Enhancer (with betaine). The patented PCR Enhancement Technology* results in many benefits for PCR. These benefits include reduced dependence of the DNA melting temperature on basepair composition, increased enzyme thermostability, suppressed DNA pausing, and decreased DNA secondary structure. The net result of these effects is increased dependability and consistency of PCR experiments.

The PCR Enhancer Technology is also Available for Use with Other Enzymes

If you can't use FailSafe™ or MasterAmp™ Extra-Long PCR Systems because your application or protocol specifies that you must use Taq, Tfl, Tth, Pfu, or another DNA polymerase, you can still benefit from Epicentre®'s patented PCR Enhancer Technology. Epicentre® also offers 2 kinds of MasterAmp PCR Optimisation Kits with proprietary sets of PCR PreMixes, one of which will significantly improve the PCR performance with your thermostable DNA polymerase. And, if you can't use the PreMixes in these kits because you need to use your own PCR reaction buffer, the MasterAmp™ PCR Enhancer (with betaine) is also available separately, so you can optimise your own PCR reactions.

Other Thermostable DNA Polymerases are also Available from Epicentre

Besides the FailSafe™ and MasterAmp™ Extra-Long PCR Enzyme Mixes, Epicentre® also offers Taq, Tfl, Tth, and AmpliTherm DNA Polymerases. Some of the properties of these enzymes and enzyme mixes are given in the chart below. However, although MasterAmp™ PCR Optimisation Kits or MasterAmp™ PCR Enhancer will noticeably improve amplification of a DNA template using these or other enzymes, the best results will be achieved in most cases only by using the FailSafe™ PCR System or the MasterAmp™ Extra-Long PCR Kit.

Contact Epicentre® for more information or for help in choosing the best products for your PCR or RT-PCR applications.

Epicentre®'s Technical Consultants have extensive experience in PCR and would be pleased to work with you to find the best solutions for your project. Contact them at:
Tel: (+001) 608 258 3080 or email: techhelp@epicentre.com

Last modified: 2008-05-14 16:46:46