Method for extracting bacterium genomic DNA by using magnetic nanoparticles

A magnetic nanoparticle and genome technology, applied in the field of nanomaterials and molecular biology, can solve the problems of long time consumption, low extraction efficiency, and use of toxic reagents, and achieve the effect of short time consumption, high extraction efficiency and simple operation

Inactive Publication Date: 2011-07-13
JIANGNAN UNIV
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Problems solved by technology

[0004] At present, the extraction methods widely used in bacterial genomic DNA are mainly traditional nucleic acid separation methods, which have disadvantages such as the use of toxic reagents (phenol, chloroform), long time consumption, and low extraction efficiency. , convenience requirements

Method used

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  • Method for extracting bacterium genomic DNA by using magnetic nanoparticles
  • Method for extracting bacterium genomic DNA by using magnetic nanoparticles
  • Method for extracting bacterium genomic DNA by using magnetic nanoparticles

Examples

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Embodiment Construction

[0030] (1) Preparation of magnetic nanoparticle aggregates:

[0031] ①

[0032] Weigh 0.65g of anhydrous ferric trichloride and 0.4g of trisodium citrate, measure 20ml of ethylene glycol, and stir for 20min to dissolve them completely;

[0033] ② Weigh 1.2g of anhydrous sodium acetate and add it to the above reaction system, stir for 10min, put it in a tetrafluoroethylene reactor and heat it to 230°C in an oil bath, and at the same time carry out magnetic stirring, stop heating after 10h of reaction, continue magnetic stirring to cool to room temperature ;

[0034] ③Add 20ml of absolute ethanol to the reacted solution, ultrasonically clean it for 10min, centrifuge at 5000rpm for 10min, and discard the supernatant;

[0035] ④ Add 20ml of absolute ethanol to the lower layer, ultrasonically clean for 10min, centrifuge at 5000rpm for 10min, and discard the supernatant;

[0036] ⑤ Add 20ml of deionized water to the lower sediment, ultrasonically clean for 10min, centrifuge at 50...

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Abstract

The invention discloses a method for extracting bacterium genomic DNA by using magnetic nanoparticles, belonging to the technical field of nanomaterials and molecular biology. The method comprises the steps of: preparing a magnetic nanoparticle aggregate, preparing a bacterium lysing solution, preparing and adding an absorption buffer solution, absorbing with a magnet, rinsing with 70 percent alcohol, and eluting with a TE buffer solution to obtain a bacterium genomic DNA solution. Compared with the traditional phenol/chloroform DNA extracting method, the method for extracting the bacterium genomic DNA by using magnetic nanoparticles has the advantages of short experiment time, simpleness in operation, high extraction efficiency, rapidness, convenience, sensitivity, high yield and the like; and the extracted bacterium genomic DNA can be completely used for subsequent experiments, such as DNA hybridization, PCR (Polymerase Chain Reaction), and the like, and provides the basis for the subsequent research and analysis.

Description

technical field [0001] The invention relates to a method for extracting bacterial genome DNA by using magnetic nanoparticles, belonging to the technical fields of nanomaterials and molecular biology. Background technique [0002] Bacteria are widely distributed in nature, and some pathogenic microorganisms can invade human body and animals, cause infection or even infectious disease, and pose great harm to human and animal health. Eating food contaminated with these bacteria and drinking water contaminated with bacteria are the main routes of transmission of foodborne diseases. According to statistics, at least 200,000 to 400,000 people actually suffer from food poisoning every year in my country. In recent years, food poisoning has increased year by year. There are many pathogenic microorganisms that cause food poisoning, the most common being Vibrio enteritidis, Salmonella, Campylobacter and pathogenic coliform bacteria. [0003] The traditional method of bacterial dete...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12N15/31
Inventor 王利兵胥传来勇倩倩邓小芳屈昌龙刘微波赵书阁
Owner JIANGNAN UNIV
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