Taq DNA Polymerase
(Concentration 5 u/μl)

Source: Purified from an E. coli strain carrying a plasmid with Taq DNA polymerase gene from Thermus aquaticus YT-1.

Description: Taq DNA Polymerase is a thermostable enzyme that catalyzes 5'→3' synthesis of DNA. The enzyme has no detectable 3'→5' proofreading exonuclease activity, but possesses low 5'→3' exonuclease activity.

Unit definition: One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmoles of dNTPs into acid insoluble material in 30 minutes at 72^oC.

Reaction conditions: 1x Taq polymerase buffer [50 mM KCl, 10 mM Tris-HCl pH 8.5 @ 25^oC, 1.5 mM MgCl₂, 0.1% Triton X-100].

Quality Control Assays:

•     Standard DNA Polymerase Assay Conditions (not PCR conditions): The polymerase activity is assayed in 10 mM Tris-HCl (pH 8.5), 50 mM KCl, 1.5 mM MgCl₂, 200 μM each of dATP, dGTP, dCTP, dTTP (a mix of unlabeled and [³H] dTTP) and 12.5μg activated calf thymus DNA, in a final volume of 50 μl.

• Functional Assay: Taq DNA Polymerase is tested for performance in the polymerase chain reaction (PCR) using 1.5 units of enzyme to amplify a 1730-bp region of the PspP I methyltransferase gene from 5 ng of bacterial genomic DNA. The resulting PCR product is visualized as a single band on an ethidium bromide-stained agarose gel.

• Absence of contaminants: Tested extensively for the absence of endo- and exodeoxyribonucleases.

Guaranteed stability: Taq DNA polymerase is guaranteed to maintain stability for six months from the date of shipment when stored as directed.

Storage Buffer: 100 mM NaCl, 50 mM Tris-HCl (pH 8.0 @ 25^oC), 1 mM DTT, 0.1 mM EDTA, 1% Triton X-100 and 50%  glycerol. Store at -20^oC.

Reagents supplied: 10x Taq DNA polymerase Buffer (1.5ml) and/or 10x Taq DNA polymerase w/o MgCl₂ (1.5ml) and 25mM MgCl₂ (1.5ml).