Paq5000 DNA Polymerases & PCR Master Mixes - Details & Specifications

Paq5000 Hotstart DNA polymerase is a robust, economic alternative to Taq DNA polymerases utilizing hot start technologies. It has been optimized to provide improved hot start capability, higher specificity, and better PCR yields with reduced cycling time. The Paq5000 Hotstart DNA polymerase employs a unique built-in hot start method developed internally by our scientists and can be easily substituted into standard PCR protocols with minimal optimization.

Amplification of a 3.9 kb fragment of the Human a1 Antitrypsin gene using Paq5000 Hotstart DNA polymerase and hot start Taq polymerases from various suppliers. All reactions were performed using each manufacturer’s recommended buffer, enzyme concentration and cycling conditions (polymerases I and C are antibody hot start, polymerases Q, R, and A are chemical hot start).

Breakthrough Hot Start Technology

Hot start methods for PCR prevent mispriming events, improve PCR yields, and allow room-temperature reaction setup. Our novel hot start method utilizes a heat-sensitive blocking polymerase protein that has been mutated to inactivate catalytic activity while retaining DNA binding activity. At room temperature, this protein binds to primed templates in the PCR reaction and prevents extension by the Paq5000 DNA polymerase. The blocking protein is inactivated during the initial PCR denaturation step, allowing the Paq5000 DNA polymerase to access the templates properly primed during the annealing step and proceed with extension.

Unlike traditional chemical or antibody methods, our method achieves true hot start without any modification to the DNA polymerase. Since we utilize the concept of competitive binding to block the DNA polymerase from extending at room temperature, the DNA polymerase is not modified to prevent extension. This allows the full activity of the DNA polymerase once the blocking protein is inactivated, leading to higher PCR yields. In addition, our method does not require a lengthy heat inactivation step that can compromise the polymerase and template DNA, nor an expensive antibody that must be used in vast excess and can interfere with the PCR reaction.

Unprecedented Pricing for a DNA Polymerase with Hot Start Capability

Our internally developed technology allows us to offer the first truly low cost hot start DNA polymerase. You can now use a hot start polymerase for the same price as a Taq DNA polymerase without hot start capability. The economic pricing makes the Paq5000 Hotstart DNA polymerase ideal for routine PCR and screening.

Faster Protocol

The combination of a quick 2 minute hot start inactivation time and just 30 seconds per kb extension time leads to a nearly 40% time savings in overall PCR run time for a 1 kb target when using Paq5000 Hotstart DNA polymerase. This allows you to save valuable time and increase throughput.

For Research Use Only. Not for use in diagnostic procedures