The FastGene® Optima is a mixture of a highly purified Taq Polymerase and a modified type-B polymerase with proof-reading abilities. The enzymes are purified using three different chromatography technologies and result in a very high purity and very high activity. The FastGene® Optima is extremly robust and was developed for the standard PCR, difficult as well as very long amplicons of over 7.5 kBp.
The FastGene® Optima can handle very complicated templates. The highly purified Taq polymerase gives high efficiency while the proof-reading polymerase guarantees the fidelity. The robustness of both enzymes makes the amplification of complex tissue, such as liver (Fig. 1), possible.
Fig.1: Comparison between (A) Competitor T‘s and (B) FastGene® Optima polymerase mixture using the hard to amplify catshark liver DNA as template. The PCR product has a size of 1030 bp and was separated onto an 1.2% agarose gel. The FastGene® Optima produces much less primer dimers and has a higher amplification efficiency.
Most polymerases have a very low amplification efficiency if the template DNA is GC-rich. As seen in Fig. 2, the FastGene® Optima has an excellent amplification efficiency even with GC-rich templates, which is even higher compared to the efficiency of polymerases especially designed for GC-rich templates (Fig. 2).
Fig. 2: Comparing the ability of Competitor T‘s and FastGene® Optima polymerase mixture to amplify GC-Rich DNA fragments. Two fragments of 60.7 % and 64.3% were amplified resulting in two products of 1839 bp and 1260 bp, repectively. FastGene® Optima had a higher efficiency compared to Competitor T‘s polymerase mixture.
Data was kindly provided by Ms. Ryoko Nakayama, Department of Pathology, Tsurumi University, Japan.
The detection of single nucleotide polymorphism (SNP) requires extreme fidelity. The proof-reading activity guarantees this needed fidelity (see Fig.3).
Fig. 3: SNP typing of the ALDH gene using FastGene® Optima polymerase. The ALDH classified as human sensitivity to alcohol gene was analysed for presence of a SNP by digesting the amplification of homo- and heterozygotes using MboII.
Data was kindly provided by Dr. Che Xiano-Fang, Department of Biochemistry, Tokyo Medical University, Japan.
|LS28||FastGene® Optima PCR kit with dNTP||250 Units|
The error rate is around 10-15 times lower compared to a regular TAQ. The FastGene Optima PCR system is a blend of Taq DNA polymerase and an engineered archaeal (B-family) DNA polymerase. Both enzymes possess 5’ – 3’ polymerase activity, but only Taq possesses 5’ – 3’ exonuclease activity, and only the B-family DNA polymerase possesses 3’ – 5’ exonuclease activity.
Yes there is a 3′ A overhang.