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Thermo Scientific Phire Hot Start II DNA Polymerase is a novel PCR enzyme for routine and high throughput PCR applications. It outperforms every Taq-based hot start polymerase on the market. Phire Hot Start II DNA Polymerase is significantly faster, extremely robust, and also capable of amplifying long DNA fragments with high yields. These features are achieved through advanced protein engineering of the polymerase. It incorporates a unique double-stranded DNA binding domain which allows short extension times (10 to 15 s/kb), improves yields, and increases fidelity 2-fold compared to Taq DNA polymerase.
The hot start modification of Phire Hot Start II DNA Polymerase is based on the same Affibody inactivation technology utilized by Phusion Hot Start II DNA Polymerase. This technology increases the specificity of the PCR reaction with no additional time required for initial activation of the enzyme.
New Phire Green format is a combination of Phire Hot Start II DNA Polymerase and 5X Phire Green Buffer. The buffer includes a density reagent and two tracking dyes for direct loading of PCR products on a gel. The colored buffer does not interfere with enzyme performance and is compatible with downstream applications such as DNA sequencing, ligation and restriction digestion.
Phire Hot Start II DNA Polymerase
Phire Green Hot Start II DNA Polymerase
A 1.5 kb fragment from the human cathepsin K gene was amplified with five different hot start DNA polymerases. Phire Hot Start II DNA Polymerase amplified high amounts of specific PCR product in just 29 minutes. In contrast, the PCR protocols for hot start Taq DNA polymerases from four major suppliers (A through D) were substantially longer and resulted in lower product yields.
Five genomic DNA fragments of different lengths were amplified with three different hot start DNA polymerases. Phire Hot Start II DNA Polymerase produced all five amplicons with very high yields. The competing hot start Taq DNA polymerases produced significantly lower yields and failed to amplify the 7.5 kb fragment.1 – 0.6 kb fragment of human glutathione peroxidase 3 gene2 – 1.0 kb fragment of human glutathione peroxidase 3 gene3 – 1.5 kb fragment of human cathepsin K gene4 – 2.7 kb fragment of human Β-2-microglobulin gene 5 to 7.5 kb fragment of human Β-globin gene
A 600 bp fragment from human genomic DNA was amplified with five different hot start DNA polymerases. With Phire Hot Start II DNA Polymerase, the PCR protocol was completed up to four times faster than with Taq DNA polymerases utilizing chemically modified or antibody-based hot start technologies (suppliers A through D).
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