Estabelecimento de protocolos de extração para araçazeiro vermelho (Psidium cattleianum Sabine) visando estudos moleculares

Abstract

Among all the genera of the Myrtaceae family, Psidium can be highlighted as one of the holders of species with the greatest commercial value, which includes araçazeiros. Research on the genetic improvement of species is important for modern agriculture, developing superior cultivars, with greater productivity and better adaptation. Currently, methods for extracting nucleic acids that are fast, low cost, with products free of contamination and toxicity, as well as efficiency, even with a small amount or scarcity of the sample, are being researched. Therefore, the objective of the present study was to evaluate the effectiveness of three DNA extraction protocols with the Wizard® Kit from Promega, for Araçá Vermelho (Psidium cattleianum SABINE), two teachers for lyophilized and one for fresh leaves. The experimente was carried out in the Molecular Biology laboratory of the Federal Technological University of Paraná - Câmpus Dois Vizinhos, PR. The ten collected at the UTFPR-DV experimental farm and the other ten collected in the municipality of Verê-PR, were subjected to lyophilization and then two protocols for lyophilized leaves were tested, one of which using proteinase K with different incubation times in water bath; thirty and sixty minutes. The other protocol uses natural leaves collected from ten trees at Câmpus Dois Vizinhos and taken immediately to the laboratory, where they were macerated with the aid of a crucible and subjected to extraction. The three methods were positive. Being the most suitable or natural extraction because it requires less time in execution. The protocol that indicates the best quality and the most time consuming was that of freeze-dried leaves. The method in which a proteinase K is used is indicated for cases where the presenters present some contamination and extraction is more difficult.