About the Figure: Using a fast polymerase to detect influenza. Scientists at Osaka University developed a microfluidic device for rapid detection of influenza A (shown in color in the figure above). Using a mix of Takara® PrimeScript™ RTase, PrimeSTAR® Max polymerase and SpeedSTAR™ polymerase, RT-PCR was complete in 15 min. Yamanaka, K., et al. (2011) Analyst 136:2064.
A fast polymerase for high fidelity PCR- save your sanity as well as your time
Let's face it: there's nothing more frustrating than failed PCR. When all that stands between you and understanding your gene of interest is DNA amplification, every moment counts. Rely on PrimeSTAR Max DNA Polymerase for cloning and expression, structure-function, and genetic variation studies.
Experimental Example: PrimeSTAR Max vs. Other Commercially Available High-Fidelity Polymerases Using a 10-second Extension Step
Figure 1. Fast PCR amplification of human genomic DNA using PrimeSTAR Max DNA Polymerase or high-fidelity polymerases provided by other manufacturers. For full experimental details, download the Application Note.
|Human genomic DNA was used as template for fast PCR. Products 0.5 to 7.5 kb long were amplified (extension time, 10 sec.) using PrimeSTAR Max enzyme or polymerases from three other manufacturers. High yield was observed when PrimeSTAR Max polymerase was used for products up to 4 kb in length.|
Compare Extension Step Recommendations
Is every high-fidelity polymerase capable of fast extension? No. Many high-fidelity polymerases on the market lack sufficient extension speed and are incompatible with fast cycling conditions. Refer to the table below for recommended extension step times for PrimeSTAR Max DNA Polymerase and three other commercially available high-fidelity enzymes.
|Polymerase||Total extension step time per cycle for a 4 kb product|
|PrimeSTAR Max polymerase||10 sec. minimum*|
|Company T polymerase||120 sec.|
|Company N polymerase||120 sec.|
|Company I polymerase||240 sec.|
*See data in Figure 1 for an example of amplification of a 4-kb product with a 10 sec. extension step. Routine recommendations: when template quantity is 200 ng/50 µl or less, an extension step of 5 sec./kb is recommended. When template quantity exceeds 200 ng/50 µl, an extension step of 30-60 sec./kb is recommended.