Direct PCR to genotyping – You’re covered.
Optima Polymerase is actually a mixture of two different types of PCR enzymes – a Taq polymerase and a modified type-B polymerase with excellent proof-reading abilities. Each enzyme is purified using three different chromatography technologies which results in a very high purity and very high activity. Optima is extremely robust making it ideal for a broad range of PCR applications. Standard PCR, difficult PCR, and very long amplicons (over 7.5 Kb) are a piece of cake for this enzyme mixture.
For those labs that prefer low primer-dimer and easy, room temperature set-up, then the HotStart-version of Optima is your best choice. Designed as a master mix, Optima HotStart ReadyMix combines the superb efficiency and robustness of the Optima enzyme with a proprietary antibody that inhibits preliminary unspecific reaction. This antibody is permanently denatured during the primary activation step. The HotStart ReadyMix comes with all the necessary ingredients for optimal PCR. Just add your template and primers. Additionally, the ReadyMix contains a loading dye, meaning that the PCR sample can be directly loaded onto an agarose gel.
Optima robustness for very complex samples
The Comparison between (A) the “best-selling” blended Taq mix 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.
Optima for SNP-typing
The detection of single nucleotide polymorphism (SNP) requires extreme fidelity. The proof-reading activity guarantees this needed fidelity. To the right, SNP typing of the ALDH gene using FastGene Optima polymerase. The ALDH classified as human sensitivity to alcohol gene was analyzed 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.
Direct PCR from Escherichia coli colonies
Below is an example of direct PCR from E. coli colonies using FastGene Taq Optima HotStart ReadyMix (left gel) or “best-selling” blended Taq mix (right gel). The ReadyMixes were used to determine the presence or absence of inserts. The Optima HotStart ReadyMix yielded a clear electrophoretic pattern without smearing. In addition, Optima was able to amplify clean product from EVERY colony. The competitor was not able to amplify 10 colonies.
Optima PCR Products
|Contents||500 x 25 reactions of 2X Master Mix:
FastGene Optima Buffer (1X)
dNTPs (0.3 mM of each dNTP at 1X)
MgCl2 (2.0 mM at 1X) and stabilizers
|2X Master Mix||Yes|
|Fidelity||2 to 4 fold higher than native Taq DNA|
|Processivity||Similar to native Taq|
|Amplicon Length||very long, over 7.5kb|
|Storage||1 year at -20°C|