Optimization of soybean dna extraction under different storage and development periods

Abstract

DNA extraction of plants with high quality is very important to researches in molecular biology.Several extraction protocols have been used to obtain soybean DNA; however, there is a lack of papers about extraction protocols optimization and the best developmental stage of the plant to collect them. Therefore, the main purpose of the study was to extract high quantity and quality of DNA from fresh or frozen soybean samples, using different protocols. Moreover, we analyzed the best developmental stage of the plant to do the extraction. Fresh leaves or leaves kept for two years in the ultra-freezer were submitted to the DNA extraction protocols: Haberer et al., 1996 (modified); second modification from Haberer et al., 1996; Murray & Thompson, 1980 (modified) e Doyle & Doyle, 1990 (modified). Modified protocol of Doyle & Doyle was used to value the best stage to collect the leaves to do the DNA extraction. The samples were collected in the stages of development VC, V1, V2, V3, V4 and R5. The experiments were conducted in completely randomized design with 10 samples per treatment. The data underwent variance analysis and the averages were compared by the Tukey test (p<0.05). Through Doyle &Doyle, 1990 andHaberer et al., 1996 modified protocols, for both fresh and frozen samples, it was possible to obtain a higher total DNA concentration if compared to the other tested protocols. However, the quality of DNAs extracted by the protocol Doyle & Doyle, 1990 (modified) was superior, due to a minor molecular degradation. Besides that, the extractions made with these protocols have shown to be more efficient using frozen leaves' tissue. Higher DNA concentrations were obtained analyzing VC samples; however, there were no statistical differences between the stages VC, V2 and V3. It is suggested thereby to use modified of Doyle & Doyle for DNA extraction from soybean leaves in V2 and V3 stages of development from frozen samples, providing the collect of a large number of samples and its storage until the analysis.

Preservation and maceration of amazon açai leaflet tissue to obtain genomic DNA

“…Our results corroborate those reported in the literature for some species of plants where the use of fresh, frozen, and cooled leaf tissue have been the predominant methodologies for preservation of Preservation and maceration... AZÊVEDO, H. S. F. S. et al biological material for subsequent genomic DNA extraction (SILVA et al, 2010;MESQUITA et al, 2015). In the present study, storage of leaflets in transport buffer for one week also proved to be a viable methodology for the preservation of plant tissue, since these leaflets retained the characteristics of fresh leaflets.…”