High fidelity Up to 5 kb

DESCRIPTION
AccuTherm™ is a thermostable enzyme possessing 5'-3' DNA polymerase and 3'-5' proofreading exonuclease activities. It is isolated from the hyperthermophilic marine archae Pyrococcus furiosis (Pfu). Base misinsertions that may occur during polymerization are rapidly excised by the proofreading activity of the polymerase. AccuTherm™ DNA Polymerase generates blunt-ended PCR fragments. AccuTherm™can amplify fragments up to 5 kb at extremely high fidelity. A mixture of either KlenTherm™ or BioTherm™ with AccuTherm™ provides more robust synthesis of longer amplification products (Barnes, 1994. Proc. Natl. Acad. Sci. USA 91:2216-2220).

APPLICATION
Due to its 3'-5' proofreading exonuclease activity, AccuTherm™ DNA Polymerase is recommended for use in PCR and primer extension reactions that require high-fidelity synthesis.

CONCENTRATION
5 units/µl

UNIT DEFINITION
One unit is defined as the amount of enzyme that incorporates 10 nmoles of dNTPs into acid-insoluble form in 30 minutes at 72°C under the assay conditions (25 mM TAPS (tris-(hydroxymethyl)methyl-amino-propane-sulphonic acid, sodium salt) pH 9.3 (at 25°C), 50 mM KCI, 2 mM MgCl₂, 1 mM β-mercaptoethanol) and activated calf thymus DNA as substrate.

STORAGE BUFFER
10 mM K-phosphate buffer pH 7.0, 100 mM NaCI, 0.5 mM EDTA; 1 mM DTT, 0.01% Tween 20; 50% glycerol (v/v)

STORAGE TEMPERATURE
Store AccuTherm™ polymerase below 0°C, preferably at -20°C, in a constant temperature freezer.

5X REACTION BUFFER
83 mM (NH₄)₂S0₄, 350 mM Tris-HCI pH 8.8 (at 25°C), 22 mM MgCl₂, 0.75% Triton X100, 100 mM KCl

PRODUCT CITATIONS
Fiedler, T.; Mix, M.; Meyer, U.; Mikkat, S.; Glocker, M. O.; Bahl, H. & Fischer, R.-J. (2008), J Bacteriol 190(20), 6559--6567.
Jahromi, Z. M.; Salmanian, A. H.; Rastgoo, N. & Arbabi, M. (2009), Hybridoma (Larchmt) 28(5), 305--313.
Moreno, I.; Norambuena, L.; Maturana, D.; Toro, M.; Vergara, C.; Orellana, A.; Zurita-Silva, A. & Ordenes, V. R. (2008), J Biol Chem 283(15), 9633--9641.