PCR-Based Assays for the Detection and Quantitation of DNA Damage and Repair

摘要: Exposure to genotoxic agents from both environmental and endogenous sources which result in damage cellular DNA poses a significant health risk the individual if such is left unrepaired. Several human diseases, including Cockayne’s syndrome xeroderma pigmentosum, have been associated with defects repair of (Friedberg et al., 1995). An overall decrease capacity observed latter, whereas former defect transcription-coupling factor facilitates rapid transcribed strand an active gene. One general method for analysis gene- strand-specific based on Southern breaks induced by damage-specific endonuclease (Smith Mellon, 1990; Bohr Okumoto, 1988). This endonuclease-sensitive site (ESS) technique employs use T4 V cleaves at pyrimidine dimers (Ganesan 1980), eliminating its ability hybridize radioactive probe alkaline analysis. Although this methodology has pivotal elucidation gene-specific single-copy genes (Bohr 1987; Mellon Hanawalt, 1989), there are certain limitations. It requires some information regarding restriction sequence flanking gene or genomic segment interest, lesion-specific incise near damaged base, perhaps most requirement large quantities (5–10 μg) generally used assays.