Kit for extracting genome DNA (Deoxyribose Nucleic Acid) from plant leaves based on paramagnetic particle method and extracting method thereof

Abstract

The invention belongs to the technical field of molecular biology, and particularly relates to a kit for extracting genome DNA (Deoxyribose Nucleic Acid) from plant leaves based on a paramagnetic particle method and an extracting method thereof. The kit for extracting genome DNA from plant leaves based on the paramagnetic particle method comprises a pretreatment solution, a rapid cracking liquid, a DNA binding liquid, a magnetic bead suspension and an eluent, wherein cell walls and cell membranes can be effectively cracked through the pretreatment solution; and the interference of impurities such as polysaccharides, polyphenol, tannin and the like on genome DAN extraction can be effectively eliminated through the pretreatment solution. In the rapid cracking liquid, guanidine hydrochloride and Tween 20 are taken as main components, so that plant cells can be fully cracked at the normal temperature within 2 minutes, only two minutes are required in a subsequent centrifuging step, and the operation time is greatly saved. The guanidine hydrochloride is a strong denaturant, has a good cell cracking effect. Moreover, organic solvents such as chloroform and the like are not required to be added, so that damages to operating personnel are avoided, and safety and reliability are achieved.

Description

Technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a kit for extracting genomic DNA from plant leaves based on a magnetic bead method and an extraction method thereof. Background technique [0002] With the continuous development of molecular biology and biomedicine, genomic DNA has become an important research topic. Genomic DNA plays a controlling role in biological growth, inheritance, mutation and other life processes. The study of genomic DNA first involves the extraction of genomic DNA. [0003] For plants, the main steps for extracting genomic DNA are: wall breaking, membrane breaking, impurity removal, rinsing and elution. First, during the extraction process, the use of decontamination reagents such as SDS or CTAB can destroy the cell membrane, precipitate intracellular proteins, and thereby release intracellular genomic DNA; secondly, use high-salt precipitation methods or chloroform-phenol extrac...

AI Technical Summary

Problems solved by technology

[0021] 1. The versatility is not strong

It works well for extracting genomic DNA of ordinary plants such as rice and wheat, but for extracting genomic DNA of plant leaf tissues rich in polysaccharides, polyphenols, tannins, lipids and other secondary metabolites such as cotton, grapes, and blackcurrant , its purification success rate is low, polyphenols and polyphenols cannot be completely removed

During the extraction process, polyphenols will be oxidized to cause browning, and polysaccharides and other substances will combine with genomic DNA to produce a sticky jelly. Both of these situations are likely to cause genomic DNA to degrade, resulting in poor purity of the extracted genomic DNA. high

[0022] 2. The lysate needs to be heated in a 60°C water bath for 30 minutes and centrifuged for 10 minutes to lyse and grind the leaf tissue. These steps consume too much time

[0023] 3. When extracting the genomic DNA of plant leaf tissue rich in secondary metabolites such as cotton, it is necessary to add harmful organic solvents such as chloroform, which is not good for the health of operators, and the waste liquid produced at the same time will also cause pollution

Images