Application of the Arbitrarily Primed Polymerase Chain Reaction for the Detection of DNA Damage

摘要

The technique of arbitrarily primed polymerase chain reaction (AP-PCR) shows potential as a selective and sensitive assay for the detection of xenobiotic-induced DNA damage. Problems, however, may occur in AP-PCR, diminishing its dis- Criminative abilities. These problems, include the presence of spurious amplifica- tion products in non-template-containing negative control reactions, and a lack of reproducibility amongst amplification patterns. Experiments designed to remove contaminated nucleic acids by ultraviolet (UV) treatment indicated that spurious bands are the result of aberrant primer-induced polymerization, an event showne to be influenced by he concentration of deoxynucleotide triphosphates (dNTP) present in the reaction mixtures.