Use of UV irradiation to reduce false positivity in polymerase chain reaction.

Abstract

UV irradiation provides a simple and efficient way to minimize contamination or false positivity which often occurs in laboratories performing routine PCR tests. Here, we characterize several parameters of the effect of UV irradiation on DNA template, primers, deoxynucleoside triphosphate and Taq polymerase. UV irradiation of DNA results in the formation of pyrimidine dimers and thus prevents them from being effective templates in subsequent PCR. Reduction of the HIV DNA templates in polypropylene microcentrifuge tubes by more than 1000-fold can be achieved by UV irradiation. The sensitivity of the primers is sequence- and concentration-dependent. Oligonucleotides with neighboring thymine bases are more susceptible to UV than those without. Taq polymerase is highly UV sensitive, whereas deoxynucleotide triphosphate is relatively UV resistant.