Phusion™ Hot Start II DNA Polymerase & dNTP Mix (10 mM each)

With Phusion Hot Start II High-Fidelity DNA Polymerase, amplification proceeds without the production of nonspecific products due to the combination of Phusion DNA Polymerase and a reversibly bound, specific Affibody ligand that inhibits DNA polymerase activity at room temperature. The Affibody ligand also inhibits the 3' to 5' exonuclease activity of the polymerase, thus preventing degradation of primers and template DNA during reaction setup. At temperatures that promote polymerase activity, the ligand is released, rendering the polymerase fully active. Phusion Hot Start II High-Fidelity DNA Polymerase is immediately reactivated at high temperatures and does not require a separate activation step in PCR protocols.

Features

• High fidelity (52X Taq)
• No non-specific amplification and primer degradation during reaction setup
• Fast PCR due to short extension times (15–30 s/kb)
• Increased product yields with minimal enzyme amounts

Applications

• Standard PCR
• qPCR
• High fidelity and long PCR
• LAMP-PCR
• cDNA synthesis
• RT-PCR, RDA, MDA
• DNA labeling and sequencing

dNTP Mix

Thermo Scientific dNTP Mix (Cat. No. R0192) contains pre-mixed aqueous solutions of dATP, dCTP, dGTP, and dTTP, each at a final concentration of 10 mM. The nucleotides have greater than 99% purity and are free of nuclease activities, as well as human and E. coli DNA. Mixes offer the possibility of reducing the number of pipetting steps and the risk of reaction setup errors.

• Greater than 99% purity confirmed by HPLC
• Free of human and E. coli DNA
• Stable after multiple freeze-thaw cycles
• Up to 95% of dNTPs remain in triphosphate form after 7 weeks at room temperature
• Up to 90% of dNTPs remain in triphosphate form after 30 cycles of PCR (1 minute at 94°C; 3 minutes at 72°C)

Notes

• This package includes Phusion Hot Start II High-Fidelity DNA Polymerase (4 x 500 U) and high-quality dNTP Mix (10 mM each, 2 x 1 mL).
• Annealing rules for Phusion DNA polymerases are different from many common DNA polymerases (such as Taq DNA polymerases).