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

A plant leaf and extraction method technology, applied in the field of molecular biology, can solve the problems of inability to completely remove polyphenols, low purification success rate, waste liquid pollution, etc., and achieve the effects of easy automation, high throughput and high purity

Inactive Publication Date: 2012-08-01
苏州启巧生物科技有限公司
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  • Abstract
  • Description
  • Claims
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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 thes

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

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Embodiment Construction

[0060] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention, and equivalent replacements in different forms are possible. This embodiment cannot limit the protection scope of the present invention.

[0061] 1. Kit

[0062] A kit for extracting genomic DNA from plant leaves based on a magnetic bead method, comprising: a pretreatment solution, a rapid lysate, a DNA binding solution, a magnetic bead suspension and an eluent,

[0063] The pretreatment solution includes: 0.2M tris-HCL, 0.05M EDTA, 2% mercaptoethanol by volume and 2% PVP by mass / volume ratio,

[0064] Rapid lysate includes: 100mmol / L KAC, PH=5.2, 50mmol / L EDTA, 6MGHCl, 5mM sodium citrate, 2% Tween 20,

[0065] DNA binding solution includes: 5% PEG8000, 4M GITC, 0.5M NaCl,

[0066] Magnetic bead suspension includes: magnetic beads dissolved in ultrapure water, wherein the conce...

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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...

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Application Information

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IPC IPC(8): C12N15/10
Inventor 李宗飞
Owner 苏州启巧生物科技有限公司
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