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Reagent and method capable of simply and effectively extracting plant pathogenic fungus genomic DNA

A plant pathogenic fungi and genome technology, applied in the biological field, can solve the problems of long consumption time, cumbersome process, and many impurities, and achieve the effects of good integrity and stability, simple and effective extraction, and high-efficiency extraction.

Active Publication Date: 2019-09-06
SOUTHWEST UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because fungi have a hard cell wall structure and contain various complex polysaccharides, the process of extracting fungal genomic DNA is generally cumbersome and contains many impurities.
[0004] The Chinese patent with publication number CN109666671A discloses a method for extracting plant pathogenic fungus DNA, which is simple and fast, but this method is not suitable for the extraction of mulberry pathogenic fungus genomic DNA
Moreover, the traditional DNA extraction methods such as SDS and CTAB methods are complicated in operation steps, take too long, the DNA quality is unstable, and the cost of the kit method is relatively large.

Method used

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  • Reagent and method capable of simply and effectively extracting plant pathogenic fungus genomic DNA
  • Reagent and method capable of simply and effectively extracting plant pathogenic fungus genomic DNA
  • Reagent and method capable of simply and effectively extracting plant pathogenic fungus genomic DNA

Examples

Experimental program
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Embodiment 1

[0035] A simple and effective method for extracting genomic DNA of plant pathogenic fungi, comprising the steps of:

[0036] (1) Get 0.2 g of Sclerotinia mulberry hypertrophic sclerotinia-Mulberry mulberry, grind it to powder with liquid nitrogen, and quickly transfer it to a sterilized centrifuge tube;

[0037] (2) Add 600 μL of solution A, vortex for 1-2 minutes, and let stand for 10 minutes; the formula of solution A is as follows: 200mM Tris-HCl (pH8.0), 250mM NaCl, 25mM EDTA, 0.5wt% SDS, 1wt% β-mercapto Ethanol, RNase 8 μL.

[0038] (3) Add 260 μL of solution B to the centrifuge tube in step (2), mix well, let stand for 10 minutes, and centrifuge at 12000 rpm for 5 minutes; the formula of solution B is: 5M KAC;

[0039] (4) Take the supernatant from the centrifuge tube in step (3) into a new centrifuge tube, add an equal volume of isopropanol, mix well, let stand for 2 minutes, and centrifuge at 12000 rpm for 5 minutes;

[0040] (5) Discard the supernatant in the centri...

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Abstract

The invention discloses a reagent and method capable of simply and effectively extracting plant pathogenic fungus genomic DNA. The extracting reagent comprises a solution A and a solution B, wherein the solution A comprises 200mM Tris-HCl (pH 8.0), 250mM NaCl, 25mM EDTA, 0.5wt% of SDS, 1wt% of beta-mercaptoethanol, and 8 microliter of RNA enzyme; the solution B is potassium acetate with the concentration being 5M. When the reagent is used for extraction, high-purity and good-stability genomes can be extracted fast and effectively, the extracted genomes can be directly used for gene cloning andgenome sequencing, and an efficient and feasible molecular detection approach is provided for pathogenic fungus molecular biology study.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a simple and effective method for extracting genome DNA of plant pathogenic fungi. Background technique [0002] The sericulture industry has a history of more than 5,000 years in my country and has made great contributions to the economic development of the Chinese nation and the spread of silk culture. As the fruit of the mulberry tree - mulberry, because of its rich nutritional value and medicinal value, it has been listed as a "fruit with the same origin of medicine and food", and has become a hot spot in the food processing industry in recent years. However, mulberry is vulnerable to damage by diseases and insect pests during the growth and maturity process, resulting in a decline in yield and quality. In the southwestern region, the mulberry mast bacterium-Cupella mulberry is the most harmful. Once the mulberry is infected, it will lead to a large area of ​​fruit mulberry prod...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 赵爱春张帅朱攀攀寇敏刘长英郑莎
Owner SOUTHWEST UNIVERSITY
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