Nostoc flagelliforme genome DNA extraction method

Abstract

The invention discloses a nostoc flagelliforme genome DNA extraction method. The method comprises the following steps: collecting pure culture cells of nostoc flagelliforme in a centrifugation manner and cleaning through sterile water, a 1.0% NaCl solution and sterile water in sequence; and carrying out homogenate wall-breaking; adopting the following extracting solutions to extract DNA, wherein the extracting solutions comprise 100mM Tris-HCL, 20mM EDTANa2, 1.4M NaCl, 0.2% mercaptoethanol, and 2.0% CTAB, and the pH is 8.0. Compared with a kit extraction method, the extraction method provided by the invention has the advantages as follows: the involved experiment operation is simple, a reagent is easy to obtain, the cost is low and the yield is high. The ratio of OD<260> to OD<280> of the genome DNA extracted through the extraction method provided by the invention is about 1.8, the purity is high, a dispersion phenomenon does not exist, completeness is good, and the nostoc flagelliforme genome DNA can be directly applied to PCR, gene recombination and other high-precision molecular biology experimental research.

Description

technical field [0001] The invention belongs to the basic technical field of biological research, and in particular relates to a method for extracting high-purity Nostoc hairiform genomic DNA. Background technique [0002] Nostoc flagelliforme, also known as Nostoc hairiforme, is a bacterium of the order Nostoc cyanobacteria. It is widely distributed in deserts and barren soils in my country. It is named for its black and slender, like human hair. Nostoc is rich in various nutrients, especially Nostoc exopolysaccharide and Nostoc phycocyanin, which have anti-aging, anti-tumor, hypoglycemic, anti-coagulant functions, and have high edible and medicinal value. The results of pharmacological tests conducted by the Japanese Institute of Microalgae and Toyama Medical and Pharmaceutical University show that the polysaccharides of Nostocystis have obvious antiviral activity against herpes simplex virus. [0003] Due to the particularity of the growing environment of Nostocs and the...

AI Technical Summary

Problems solved by technology

Although these methods can extract a certain amount of DNA, the yield is low and the purity is poor.

[0004] Due to the particularity of Nostoc hairiformis, simply repeating any of the above methods cannot well rule out the interference of extracellular polysaccharides, proteins, phenols and other substances

The existence of these substances brings great difficulties to the extraction of DNA and subsequent experiments

For example, extracellular polysaccharides can inhibit the biological enzyme activity of ligase and polymerase; the presence of protein will reduce the purity of DNA; the oxidation of phenolic substances can cause DNA degradation and discoloration, which will affect the subsequent experiments.

In addition, the plant genomic DNA extraction kit commonly used in the laboratory is a centrifugal column kit, which needs to pass through the adsorption column, the DNA yield is low, the purity is poor, and the cost of the sample is high

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