Anti-Taq high is a highly purified neutralization monoclonal antibody to Taq DNA polymerase. This product provides an antibody-mediated hot start PCR to enhance the specificity and sensitivity of PCR. This antibody inhibits polymerase activity before the onset of thermal cycling, preventing primer dimer formation and non-specific amplification. At the first denaturation step of the thermal cycling, the antibody is quickly inactivated and PCR proceeds. The antibody-mediated hot start method is significantly more convenient to use than other hot start methods.
- Inhibits ≥95% polymerase activity at 45ºC
- No contamination of mouse genomic DNA examined by PCR
- Enhances the specificity and sensitivity of PCR
- Can be used when other hot start methods are difficult to perform (e.g. capillary PCR)
- The polymerase can be reactivated quicker than with methods utilizing a chemically modified polymerase
- Antibody-mediated hot start PCR
10 mM Tris-HCl (pH7.0), 50 mM KCl, 50% glycerol
Store at -20ºC
Anti-Taq high (1µg/µl)*
10 x PCR buffer
*100 µl of Anti-Taq high; corresponds to 500 U Taq DNA polymerase
TYPICAL PCR REACTION SETUP
Example 1.Application of the hot start PCR using the Taq based high efficient PCR enzyme
The efficiency of anti-Taq antibodies were evaluated by the amplification of the human β-globin gene (3.6 kb).
The result indicates that anti-Taq high increases the specificity and sensitivity of the PCR compared with the control reaction and PCR mediated hot start using company A's anti-Taq antibody.
- R.T. D'Aquila, L.J. Bechtel, J.A. Videler, Eron JJ, P. Gorczyca, J.C.Kaplan, Maximizing sensitivity and specificity of PCR by pre-amplification heating. Nucleic Acids Res. 19: 3749 (1991)
- Q. Chou, M. Russell, D.E. Birch, J. Raymond, W. Bloch. Prevention of pre-PCR mis-priming and primer dimerization improves low-copy-number amplifications. Nucleic Acids Res. 20: 1717-23 (1992)
- D.E. Kellogg , I. Rybalkin, S. Chen, N. Mukhamedova, T. Vlasik, P.D. Siebert, A. Chenchik. TaqStart Antibody: "hot start" PCR facilitated by a neutralizing monoclonal antibody directed against Taq DNA polymerase. Biotechniques. 16: 1134-7 (1994)