出版社：Grupo de Pesquisa Metodologias em Ensino e Aprendizagem em Ciências
摘要：DNA-based identification tools are of great importance in plant genetic studies, where DNA isolation and purification are crucial steps. The aim of the present study was to test the efficiency of conservation and maceration methods of biological materials in the extraction of genomic DNA from leaf and stipe of x Butyagrus nabonnandii (Prosch.) Vorster. The data were analyzed with the help of the software Genes via variance analysis, considering the factorial scheme 3x2x2x2, with three replications (three specimens of the hybrid; two types of biological material – stipe and leaf; two types of conservation – fresh and dehydrated; two types of maceration – with and without liquid nitrogen). The means were analyzed by Tukey test at 5% error probability. Genomic DNA was evaluated by spectrophotometry to determine the concentration. Integrity was determined by agarose gel eletrophoresis. Two primers from the WRKY gene Family were used to test the quality of the DNA obtained in PCR reactions. Statistical analyses revealed significant differences in DNA concentrations between the evaluated methods. Higher amounts of DNA were obtained from the extraction of fresh leaves when compared to dehydrated ones. DNA yield from fresh and dehydrated stipe did not vary between the methods tested. The use of liquid nitrogen can be dispensed, as a result of band patterns in PCR reactions. It was possible to extract quality DNA and sufficient amounts of biological materials from x B. nabonnandii, which amplified the genomic regions of interest.
