A novel method of genomic DNA extraction for Cactaceae1
نویسندگان
چکیده
UNLABELLED PREMISE OF THE STUDY Genetic studies of Cactaceae can at times be impeded by difficult sampling logistics and/or high mucilage content in tissues. Simplifying sampling and DNA isolation through the use of cactus spines has not previously been investigated. • METHODS AND RESULTS Several protocols for extracting DNA from spines were tested and modified to maximize yield, amplification, and sequencing. Sampling of and extraction from spines resulted in a simplified protocol overall and complete avoidance of mucilage as compared to typical tissue extractions. Sequences from one nuclear and three plastid regions were obtained across eight genera and 20 species of cacti using DNA extracted from spines. • CONCLUSIONS Genomic DNA useful for amplification and sequencing can be obtained from cactus spines. The protocols described here are valuable for any cactus species, but are particularly useful for investigators interested in sampling living collections, extensive field sampling, and/or conservation genetic studies.
منابع مشابه
A Simple and Rapid Leaf Genomic DNA Extraction Method for Polymerase Chain Reaction Analysis
In plants, secondary metabolites and polysaccharides interfere with genomic isolation procedures and downstream reactions such as restriction enzyme analysis and gene amplification. The removal of such contaminants needs complicated and time-consuming protocols. In this study, a simple, rapid and efficient method for leaf DNA extraction was optimized. This method use small amount of plant mater...
متن کاملComparative Study of Extracted Nucleic Acid from Escherichia coli by Two Methods Phenol-chloroform and Extraction Using Magnetic Nanoparticle
Background: The most important researches in molecular and genetic engineering fields are the finding of optimal methods for extraction of the genomic content of microorganisms and cells using them, can achieve the most amount and purity with the least time and cost. There are several methods for the extraction of bacterial DNA and the use of magnetic nanoparticles is one of the novel methods o...
متن کاملDNA Extraction of Almond without Phenol and Liquid Nitrogen
Genomic DNA extraction with a high quantity and quality is a basic requirement in molecular biology. The DNA obtained was free of any contamination proteins, polysaccharide, polyphenols and colored pigments. These compounds would interfere with the genomic isolation procedures and downstream reactions such as restriction enzyme analysis and gene amplification. The isolated genomic DNA was fo...
متن کاملAn efficient and simple CTAB based method for total genomic DNA isolation from low amounts of aquatic plants leaves with a high level of secondary metabolites
An efficient DNA isolation protocol specifically modified to get pure quality DNA required for molecular studieshas been reported in this paper. Some aquatic plants (Potamogeton spp., Ceratophyllum demersum and Myriophyllum spicatum) were used for the study. The protocol developed will be useful in getting high and pure DNA. Instead of using the available DNA extraction kits, this protocol can ...
متن کاملOptimization of the genomic DNA extraction in some mosses
The presence of organic compounds and high amount of secondary metabolites (polysaccharides, phenolic component, etc.) in mosses cause difficulties in DNA extraction that are followed by problems in PCR reactions. In lower plants, various methods have been used for DNA extraction including silica gel and different commercial kits. These methods mostly use hazardous (like phenol or liquid nitrog...
متن کامل