When choosing a Taq DNA Polymerase, an investigator wants to achieve high yield and fidelity. Some enzymes, such as Pfu Polymerase, have a remarkably low mis-incorporation rate, but also a rather low yield, affecting product availability for simultaneous applications. Regular Taq Polymerase provides high yield, but is famous for its high error rate. Platinum Taq DNA Polymerase High Fidelity offers the best of both yield and fidelity and therefore, is a good choice when quantity and quality are important. Platinum Taq DNA Polymerase is composed of recombinant Taq DNA Polymerase, Pyrococcus GB-D Polymerase and Platinum Taq Antibody. Pyrococcus GB-D Polymerase possesses proofreading ability and (when mixed with regular Taq Polymerase) increases fidelity by approximately six fold. An Anti-Taq DNA Polymerase provides an automatic “hot-start” for Taq Polymerase by binding to the enzyme and inhibiting its activity until the temperature reaches 94 degrees. Hot start is known to positively affect the PCR reaction by increasing sensitivity, specificity and yield. It also provides the convenience of assembling reactions at room temperature. It is important to note that some experimental systems, such as Expressway Linear Expression System by Invitrogen, are optimized for use with Platinum Taq DNA Polymerase High Fidelity.
Platinum Taq DNA Polymerase High Fidelity comes in three different sizes: 100, 500 and 5000 reactions. The 10X High Fidelity PCR Buffer is provided in the kit, as well as, standard 50 mM magnesium sulfate solution.
In our laboratory, we have routinely used a number of different enzymes, mainly regular Taq DNA Polymerases. Whenever routine Taq Polymerase amplifications did not produce satisfactory results, we substituted AccuPrime Taq DNA Polymerase (Invitrogen). Initially, Platinum Taq DNA Polymerase High Fidelity was only used in conjunction with Expressway Linear Expression System with TOPO tools technology (Invitrogen) as suggested by manufacturer’s protocol. But after evaluating the results, we started to routinely use it instead of Taq Polymerase for demanding applications such as generating high quality high yield DNA templates for in-vitro transcription experiments. Platinum Taq DNA Polymerase High Fidelity consistently gave us band intensity equal or exceeding the results when using regular Taq Polymerase. Compared to AccuPrime Taq DNA Polymerase, the bands were stronger and no mis-priming was observed. An especially striking difference was noticed when some of the reagents were mixed at room temperature, because of the automatic “hot-start”. The high yield and robustness at different set-up conditions, together with improved error rates incorporated into the Platinum Taq DNA Polymerase High Fidelity system, satisfy our research needs. In addition, we have an advantage of being in the Invitrogen Freezer Program, which guarantees us reduced prices and immediate availability of this product.
In short, we feel that Platinum Taq DNA Polymerase High Fidelity provides a reliable performance and should be seriously considered by researchers demanding a higher performance from PCR enzymes, especially if available on site.
Yelena Bykhovskaya PhD
Research Scientist I
Molecular Hematology Laboratory
Cedars-Sinai Medical Center, Los Angeles CA