The AccuPrime™ Taq DNA Polymerase from Invitrogen is designed to prevent mispriming throughout all cycles of PCR to amplify only the specific DNA targets of interest, without non-specific bands, for accurate performance and specificity in PCR. Therefore it is great for demanding PCR applications, such as multiplex PCR, with suboptimal primer sets, or for reduction of primer-dimer in PCR. Robust performance is achieved with primer annealing temperatures with between 55°C and 65°C. The technology is based on the ability of anti-Taq DNA polymerase antibodies to inhibit enzyme activity and provide automatic hot-start. In addition, the thermostable AccuPrime™ accessory protein enhances specific primer-template hybridization during every cycle of PCR.
The AccuPrime™ Taq DNA Polymerase system provides AccuPrime™ Taq DNA Polymerase containing anti-Taq DNA polymerase antibodies, and two10X AccuPrime™ buffers I and II containing AccuPrime™ protein, Mg++, and dNTPs. Buffer I is designed for small genomic DNA amplicons (<200bp), cDNA, or plasmids. Buffer II is designed for genomic DNA (200 bp-4kb). It is a complete system, which only requires an addition of researcher-specific DNA template and primers.
We have used this system extensively in our laboratory for the last couple years whenever routine Taq Polymerase amplifications did not produce satisfactory results. Recently, we began a project which involved amplifying and sequencing more than 200 separate candidate genes. Despite of the fact that we used a computer program to design the PCR primers, in about 10% of the PCR reactions multiple bands were detected on the agarose gel after PCR using regular Taq Polymerase. For all of these reactions (where multiple bands were seen) we then used the AccuPrime™ Taq DNA Polymerase system and achieved a single PCR band of the correct size in about 90% of the reactions. The remaining 10% still contained extra PCR bands but their amount and intensity were greatly reduced. Thus, even in cases where there were more than one band, we were able to cut the correct band from the agarose gel and extract the correct PCR product for sequencing. The sizes of the PCR products varied from 300 to 1000 bp. For this application we used Buffer II supplied with the system.
We also extensively used AccuPrime™ Taq DNA Polymerase system for amplification of first-strand DNA for RT-PCR. Every time routine Taq Polymerase amplification following first-strand synthesis was not satisfactory, AccuPrime™ Taq DNA Polymerase gave a much better result. Notably, in some cases when very faint PCR product was amplified with Taq, the AccuPrime™ Taq band was much stronger. Buffer I, also supplied with the system, was used for this project. The only reason we do not routinely use it in my laboratory is because of the higher cost comparing to Taq Polymerase.
In summary, AccuPrime™ Taq DNA Polymerase system is a great product that is virtually guaranteed to give a better PCR result when Taq Polymerase fails.
Yelena Bykhovskaya, Ph.D.
Research Scientist I
Molecular Hematology Laboratory
Cedars-Sinai Medical Center