RESUMEN
In this study, we determined the optimal RAPD amplification conditions to obtain genetic molecular markers for the rapid and accurate identification of Cryptococcus spp. and Candida spp. The following parameters are modified: template DNA, DNA polymerase, magnesium cloride and primer concentration; denaturation, annealing and extension time, temperature of annealing and thermal cycles. After the optimization, reliable and reproducible RAPD patterns are obtained.
RESUMEN
RAPD analysis was used to estimate the genetic diversity in an Iberian imperial eagle (Aquila adalberti) population, one of the most threatened bird species in the world. Forty-five of 60 arbitrarily designed primers amplified 614 loci in 25 individual eagles, 59.7% of which were polymorphic. In contrast to the traditional allozyme analysis performed in a previous study, the RAPD method has revealed a high level of heterozygosity in this species (H = 0.267+/-0.008). The genetic distances estimated between 25 eagles can serve to establish more adequate mating in order to preserve genetic variability. Conservation efforts being carried out in Spain in this species might be successful based on the results obtained in the present work.
Asunto(s)
Águilas/genética , Variación Genética , Animales , Águilas/clasificación , Femenino , Masculino , Técnica del ADN Polimorfo Amplificado AleatorioRESUMEN
The mucopolysaccharide capsule of Cryptococcus neoformans and other pathogenic yeasts prevent the extraction of DNA from these important zoonotic agents. We report that the use of a lysis buffer containing a high concentration of urea is an easy, efficient and time-saving technique to obtain high yields of good-quality DNA for molecular diagnosis. The use of urea also prevents the degradation of DNA during storage of samples at room temperature for up to 6 months.