3 resultados para DNA determination

em QUB Research Portal - Research Directory and Institutional Repository for Queen's University Belfast


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Stock enhancement experiments of European lobster (Homarus gammarus) have been carried out around the Kvitsoy Islands in south-western Norway since 1990. In addition to releases of coded wire tagged lobster juveniles (cultured) and subsequent monitoring of commercial fishery, a lobster hatchery was established in 1997. Several experiments were made on the communal-rearing approach where the performance of mixed larval groups (families) was evaluated under identical conditions. Berried females of wild and cultured origin and their respective fertilised eggs were screened by using microsatellite DNA profiling involving a multiplex set of six lobster specific primers, thereby allowing determination of both parental genotypes. Each female were kept separately during hatching, and the offspring were later mixed and raised in a communal rearing system. The early-larval survival was estimated at stage IV (bottom stage), and the survivors were identified to family and group by microsatellite profiling. Five different communal experiments were conducted, representing offspring from 65 berried females. Of the surviving larvae, 6.3% could not be assigned to family due to degraded DNA and no PCR amplification. Significant differences in early survival between offspring of wild and cultured origin were found in the experiments. No differences between the groups were found in stage IV larval size. Based on the pooled data on survival (as a measure of early larvae fitness) offspring of cultured females displayed a relative fitness of 60% in comparison to offspring from wild females. Large variation in survival was also observed among families within the wild and cultured groups, suggesting a genetic component for these traits and a potential for selective breeding.

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Extremely low-frequency electromagnetic fields (ELF-EMF) have been reported to induce lesions in DNA and to enhance the mutagenicity of ionising radiation. However, the significance of these findings is uncertain because the determination of the carcinogenic potential of EMFs has largely been based on investigations of large chromosomal aberrations. Using a more sensitive method of detecting DNA damage involving microsatellite sequences, we observed that exposure of UVW human glioma cells to ELF-EMF alone at a field strength of 1 mT (50 Hz) for 12 h gave rise to 0.011 mutations/locus/cell. This was equivalent to a 3.75-fold increase in mutation induction compared with unexposed controls. Furthermore, ELF-EMF increased the mutagenic capacity of 0.3 and 3 Gy gamma-irradiation by factors of 2.6 and 2.75, respectively. These results suggest not only that ELF-EMF is mutagenic as a single agent but also that it can potentiate the mutagenicity of ionising radiation. Treatment with 0.3 Gy induced more than 10 times more mutations per unit dose than irradiation with 3 Gy, indicating hypermutability at low dose.

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The effect of a cold (<40 °C) radio frequency-driven atmospheric pressure plasma jet on plasmid DNA has been investigated. Gel electrophoresis was used to analyze the DNA forms post-treatment. The experimental data are fitted to a rate equation model that allows for quantitative determination of the rates of single and double strand break formation. The formation of double strand breaks correlates well with the atomic oxygen density. Taken with other measurements, this indicates that neutral components in the jet are effective in inducing double strand breaks.