ABSTRACT
Purified genomic DNA can be difficult to obtain from some plant species because of the presence of impurities such as polysaccharides, which are often co-extracted with DNA. In this study, we developed a fast, simple, and low-cost protocol for extracting DNA from plants containing high levels of secondary metabolites. This protocol does not require the use of volatile toxic reagents such as mercaptoethanol, chloroform, or phenol and allows the extraction of high-quality DNA from wild and cultivated tropical species.
Subject(s)
Anacardium/chemistry , Casearia/chemistry , DNA, Plant/isolation & purification , Lippia/chemistry , Liquid Phase Microextraction/methods , Plant Leaves/chemistry , Anacardium/genetics , Buffers , Casearia/genetics , DNA, Plant/genetics , Electrophoresis, Agar Gel , Hydrogen-Ion Concentration , Lippia/genetics , Microsatellite Repeats , Plant Leaves/genetics , Polymerase Chain Reaction/standards , Polyphenols , Potassium Acetate , SulfitesABSTRACT
An improved and efficient protocol was developed based on the TaKaRa RNAiso Plus Kit (Code: D9108A) for isolating good-quality total RNA from the optic stalk of mud crab, Scylla paramamosain. The protocol was based on the Trizol method with modifications. The carapace overlapping the optic stalk was retained with RNA in regular protocol. In order to remove the abundant deposition correlative with the carapace which makes the isolation of RNA particularly difficult, 5M potassium acetate solution (pH = 6.0) was added before the precipitation of RNA, and the temperature of RNA deposition was also decreased to -70ºC to ensure the stabilization of RNA. Good-quality total RNA from the optic stalk of S. paramamosain could be easily isolated with this modified protocol and three conventional methods were also employed to confirm the quality of RNA. This improved method would be helpful in facilitating molecular research of crabs involving RNA from the optic stalk.
Subject(s)
Animals , Brachyura , Optic Nerve , RNA , Cloning, Molecular , Electrophoresis, Agar Gel , Potassium Acetate , Reagent Kits, Diagnostic , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
Este trabalho objetivou avaliar os efeitos da adubação com N e K no desenvolvimento de mudas de maracujazeiro-amarelo (Passiflora edulis) cultivadas em Latossolo Vermelho distrófico. O delineamento experimental foi o inteiramente casualizado, com três repetições, num esquema fatorial 4 x 4, com 4 doses de N (zero, 150, 300 e 600mg de N dm-3) e quatro de K (zero, 75, 150 e 300mg de K dm-3), parceladas em quatro vezes. A unidade experimental foi constituída por vasos de 3,3L. Após 84 dias da germinação, foram avaliados: altura da planta, o diâmetro do caule, o número de folhas e a área foliar. O melhor desenvolvimento das mudas de maracujazeiro ocorreu com as doses de 150mg de N dm-3 e de 300mg de K dm-3.