Method for extracting DNA from dry apricot leaf

Abstract

The invention belongs to the technical field of preparation of plant DNA and relates to a method for extracting DNA from a dry apricot leaf. The method comprises the following steps: firstly extracting secondary metabolite substances from the leaf; putting the grinded dry apricot leaf in an ice bath by using a precooling buffer extract solution, centrifuging, removing the supernatant liquid, and repeatedly extracting for a plurality of times to remove the secondary metabolite substances such as polysaccharide, polyphenol and ester from the apricot leaf before the DNA is released so as to prevent the secondary metabolite substances from forming a compound with the DNA; during liquid nitrogen grinding and treatment of the buffer extraction solution, adding a proper amount of PVP-40T to protect the DNA from oxidation and promote cell disruption so that more DNA can be released; adding a proper amount of RNA enzyme after the cell disruption so that RNA is removed and the introduced protease is removed as well; and extracting by using a proper amount of a mixed solution of phenol, chloroform and isoamylol by adopting an improved CTAB method to remove a great amount of protein so that the purity and quality of the DNA in the apricot leaf are improved. Compared with a generally-used method, the method disclosed by the invention is low in cost.

Description

technical field [0001] The invention belongs to the technical field of plant total DNA preparation, in particular to a method for extracting DNA from dried apricot leaves. technical background [0002] Plant leaves collected in the wild have to be transported and preserved over long distances. Regardless of young leaves or mature leaves, after drying through silica gel, although the leaves lose water rapidly, the plant leaf tissues still contain a large amount of polysaccharides, polyphenols, and proteins. and esters and other plant secondary metabolites and degraded DNA, which makes the yield, quality and difficulty of extracting DNA very different from those extracted from fresh leaf materials. [0003] DNA is the carrier of genetic information and the basic genetic material of plants. High-quality DNA samples are the material basis for molecular biology research such as restriction enzyme digestion, PCR amplification, molecular hybridization, genetic polymorphism analysi...

AI Technical Summary

Problems solved by technology

The types and contents of secondary metabolites in plants vary greatly, and sometimes the types and contents of secondary metabolites in different states of the same plant organ or tissue are also different. The DNA extraction method optimized for a certain species may not be applicable to other species, so it is difficult to obtain DNA from high-quality apricot leaves

The traditional CTAB method is currently the most widely used DNA extraction method, but when this method extracts the dried leaves of apricot plants, a large amount of protein and sugar substances remain, which affects the subsequent PCR amplification and genetic polymorphism analysis.

So far, there have been no research reports on the extraction of high-quality DNA from dried apricot leaves

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