Taq DNA Polymerase

Source: E. coli strain expressing a full length, unmodified clone of Taq DNA Polymerase from Thermus aquaticus^(1-3).

Applications:

• Routine PCR amplification
• Suitable for real time qPCR

Description:
Taq DNA Polymerase is a thermostable enzyme which has a highly processive 5'3' polymerase activity and a 5'3' exonuclease activity. USB Taq is functionally tested and has no detectable contaminating endonucleases or exonucleases. Taq DNA Polymerase withstands repeated incubations at 95°C without a significant decrease in enzyme activity, and is suitable for routine PCR. USB Taq DNA Polymerase is supplied with 10X PCR Buffer plus a separate tube of 25mM MgCl₂ for optimizing PCR conditions. Each lot is tested for product yield and length in PCR amplification.

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Fig. 1. DNA fragments ranging from 1 kb to 6 kb amplified from λ DNA.

Properties:
Activator: Mg²⁺

Purity:
Free from detectable non-specific nucleases.

Storage Buffer:
20mM Tris-HCl, pH 8.5, 1mM DTT, 0.1mM EDTA, 100mM KCI, 50% glycerol, stabilizers.

Unit Definition:
One unit incorporates 10 nmol of total nucleotides into acid-insoluble material in 30 min at
74°C in a total volume of 50 µl.

Concentration:
5 units/µl

Assay Conditions:
The reaction mixture (50 µl) contains 25mM TAPS, pH 9.3 (at 25°C), 50mM KCl, 2mM MgCl₂, 1mM 2-mercaptoethanol, 200µM dNTPs, 250 µg/ml activated salmon sperm DNA and Taq DNA Polymerase. After incubation at 74°C for 10 min, acid insoluble material is determined.

Tested User Friendly Functional Test:
Tested in a standard PCR.

Functionally Tested 10X PCR Reaction Buffer (included, PN 71165):
100mM Tris-HCl (pH 8.6), 500mM KCl, 15mM MgCl₂

Functionally Tested MgCl₂ (included, PN 71167):
25mM solution

References:

1. INNIS, M. A., MYAMBO, K. B., GELFAND, D. H. AND BROW, M. A. (1988) Proc. Natl. Acad. Sci. 85, 9436-9440.
   
2. LAWYER, F. C., STOFFEL, S., SAIKI, R. K., MYAMBO, K., DRUMMOND, R. AND GELFAND, D. H. (1989) J. Biol. Chem. 264, 6427-6437.
   
3. INNIS, M. A. AND GEFLAND, D. H. (1990) PCR Protocols: A Guide to Methods and Applications, Academic Press.

Storage:
Shipped on dry ice. Store at -20°C.