Terms and techniques: New approach to hot-start polymerase chain reaction using Taq DNA polymerase antibody

• Published in: Urologic oncology Vol. 1; no. 1; pp. 42 - 46
• Main Authors: Dahiya, Rajvir, Deng, Guoren, Chen, Katherine, Haughney, Peter C., Cunha, Gerald R., Narayan, Perinchery
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 1995
• ISSN: 1078-1439; 1873-2496
• DOI: 10.1016/1078-1439(95)00001-X

The purpose of this study was to optimize the conditions for polymerase chain reaction (PCR). The most common problem with conventional PCR is the presence of nonspecific products and primer-dimers formation, which could be due to several factors such as annealing temperature, primer concentration, and Taq DNA polymerase activity during setup of the PCR. Recently, a neutralizing monoclonal antibody (TaqStart) that blocks Taq DNA Polymerase activity has been developed. In the present study, we determined the effects of Taq DNA polymerase monoclonal antibody (TaqStart antibody) and other parameters, such as annealing temperature and oligonucleotide primer concentrations, on the specificity of PCR products. The results of these experiments suggest that TaqStart antibody inhibits nonspecific products and primer-dimers formation by blocking Taq DNA polymerase activity until the reaction components are heated in the thermal cycler (hot start). Other factors such as annealing temperature, oligonucleotide primer concentration, and magnesium concentration are equally important for specificity of PCR products.