| Description
Taq DNA Polymerase is a thermostable enzyme of approximately 94kDa isolated from Thermus aquaticus . This unmodified enzyme replicates DNA at 74¡æ and exhibits a half-life of 40munutes at 95¡æ(1,2). The enzyme catalyzes the polymerization of nucleotides into duplex DNA in the 5' ¡ú 3' direction in the presence of magnesium and also possesses a 5' ¡ú 3' exonuclease activity. Taq DNA Polymerase is recommended for use in PCR but is not recommended for use in DNA sequencing reactions.
Quality Control
1. Standard DNA Polymerase Assay Conditions [Not PCR Conditions]: 50mM Tris-HCl (pH9.0), 50mM NaCl, 10mM MgCl 2 , 200¦ÌM each of dATP, dGTP, dCTP, dTTP (a mix of unlabeled and [ 3 H]-TTP), 12.5¦Ìg activated calf thymus DNA, in a final volume of 50¦Ìl.
2. Overdigest (OD) Assay: One ¦Ìg of ¦Ë DNA is incubated with 30 units of Taq DNA Polymerase in 1 x recommended assay buffer at a final concentration of 0.6 units/¦Ìl for 16 hours at 74 ¡æ . Upon the incubation, no degraded ¦Ë DNA was detected by agarose electrophoresis.
3. Nickase Assay: One ¦Ìg of supercoiled plasmid DNA is incubated with 30 units of Taq DNA Polymerase in 1 x recommended assay buffer at a final concentration of 2 units/¦Ìl for 4 hr. at 74 ¡æ . Upon the incubation, no nicked or linearized DNA band was detected by agarose electrophoresis.
4. Thermal Stability Assay: Half life of 5 units/¦Ìl Taq DNA Polymerase in the storage buffer at 94 ¡æ is longer than 1 hour.
Features
Reliable: Compositions of the storage buffers have been optimized to assure quality performance of the enzyme under a variety of conditions.
References
• Chien, A. et al. (1976) J. Bacteriol. 127 , 1550-7.
• Kaledin, A.S. et al. (1980) Biokhimiia. 45 , 644-51.
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