The FailSafe™ PCR System from Epicentre is a kit designed for PCR troubleshooting. It includes the FailSafe™ PCR Enzyme Mix and an extensively tested set of 12 FailSafe™ PCR PreMixes, representing various PCR conditions. Along with the FailSafe™ PCR Enzyme Mix, these PreMixes contain everything you need for a successful PCR: dNTPs, buffer, and varying amounts of MgCl2 and FailSafe™ PCR Enhancer (with betaine). FailSafe™ PCR Enzyme Mix, a unique blend of thermostable enzymes that is capable of amplifying the most difficult DNA templates with extremely high sensitivity and high fidelity. Based on side-by-side experiments to determine error rates, the FailSafe™ PCR Enzyme Mix has a fidelity at least 3 times higher than Taq DNA polymerase. Notably, there is no need for "hot start". The kit can be used for a variety of applications including amplification of templates at least 20 kb in length, amplification of GC-rich templates, and multiplex PCR. PCR Enhancer Technology guarantees extremely high sensitivity and specificity for all applications including cloning, expression, and mutation analysis.
The FailSafe™ PCR PreMix selection system is supplied as a kit, which includes FailSafe™ Enzyme Mix and twelve tubes with FailSafe™ PCR 2X Premixes numbered A through L. Individual mixes contain 100 mM Tris-HCl, 100 mM KCl, 400 uM each dNTP, and varying concentrations of MgCl2 (3 mM to 7 mM) and PCR enhancer (0 to 8X). A PCR master mix containing template, primers and the FailSafe™ Enzyme Mix is aliquoted to each of the PreMixes, followed by PCR amplification. The visualization on the agarose gel clearly shows which PreMix is best for that primer/template pair combination. Conveniently, once the optimal PreMix has been determined, that PreMix can be purchased separately or in conjunction with the FailSafe™ PCR Enzyme Mix.
In my laboratory, we perform hundreds of different PCR amplifications for varying projects. Despite the fact that we use a computer program to design optimal PCR primers, it is inevitable that some PCR reactions fail. This kit is our number one choice for PCR troubleshooting when no PCR product (or a weak PCR product) has been amplified using regular Taq DNA Polymerase. In my experience, in 90-95% of the cases at least one PreMix will yield a PCR product of the correct size. Once we establish which PreMix works best, we buy it separately in a convenient and inexpensive 2.5 ml size. The PCR products are then used for a number of downstream applications such as sequencing, cloning, and mutation analysis. Also, you do not need to buy FailSafe™ PCR Enzyme Mix every time, the buffers work just as well with regular Tag DNA Polymerase.
The only drawback we have encountered with this product is when sharing protocols with collaborators in other countries. Since the conditions are optimized with the specific PreMix, we need to supply it to the labs overseas to duplicate our results.
In short, the FailSafe™ PCR PreMix selection system is a great system for PCR troubleshooting and I use it extensively in my laboratory.
Yelena Bykhovskaya, Ph.D.
Research Scientist I
Molecular Hematology Laboratory
Cedars Sinai Medical Center