β-tubulina : avaliação da amplificabilidade do DNA genômico obtidos por dois protocolos in-house da sequência conservada da via de biosíntese de fungos filamentosos produtores de aflatoxinas

Abstract

In the present study, the polymerase chain reaction (PCR) was evaluated as a molecular technique to identify the aflotoxins filamentous fungi producers. The PCR was carry out using a specific pair of primers for b-tubulin amplification, that has a high conserved sequence in filamentous fungi. It was tested two DNA extraction methods: Lee et al. and Weising et. al. both methods were efficient in DNA isolation. The results showed that using Lee et. al. method was possible to extract DNA with better quality, and the PCR runs presented the expected band for b-tubulin. These results showed that the lee et al. method and PCR is adequate for Aspergillus sp. detection. There is a perspective to improve the detection using specific primers to amplify genes related to aflotoxin biosynthesis, allowing the development of a molecular method to evaluate the risk contamination of this substance in food.