Thursday, August 15, 2013

Successful PCR Optimization with FailSafe™

We recently received a praiseworthy testimonial for FailSafe PCR System from one of our customers at the Kennedy Institute of Rheumatology, at the University of Oxford, and thought it was worth sharing. The following submission appears in its original form, and reports of successful amplification using FailSafe™ 2X PCR PreMix D for long amplicons.

At the start of my project I had to overcome the difficulty of successfully amplifying long PCR products. I had previously used a product system specifically stated to be capable of expanding long PCR products; however, this approach was unsuccessful. The number of samples which required genotyping was increasing while I spent time adjusting the parameters of the PCR protocol, and the ability to correctly genotype my samples was critical to the progress of my research.  The decision was made to optimise the components of the PCR reaction mix.

Previous work in my laboratory involved the use of pre-mixed ‘FailSafe’ PCR buffers from Epicentre, containing all of the reagents required for PCR, with the exception of the template DNA and DNA polymerase. Epicentre FailSafe buffer D had previously been successfully used to amplify long PCR products. This buffer was incorporated into the protocol for PCR amplicon of long PCR amplicon and resulted in successful amplification compared to the competitor’s product (A). This result made it possible to identify the genotype of individual samples (B). I also used Epicentre ‘FailSafe’ PCR Buffer J to successfully and reproducibly expand smaller PCR amplification products.

These FailSafe buffers resolved my genotyping issues and have been working perfectly since I began using them. I would recommend them to other researchers who would like to simplify and resolve problems with their PCR procedures.