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Method for quickly extracting genome DNA of riemerella anatipestifer, and kit

A technique of Riemerella anatipestifer and Mergerella anatipestifer, which is applied in the field of molecular biology of microorganisms, can solve the problems of lack of versatility, low genome concentration, high cost, etc., and achieves simple and feasible operation, high extraction efficiency and strong practicability. Effect

Inactive Publication Date: 2014-03-05
INST OF ANIMAL HEALTH GUANGDONG ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these methods often have their own limitations and lack of versatility. For the extraction of the Riemerella anatipestifer genome, the effect is often not ideal, such as high cost, time-consuming, low genome concentration, and inability to extract in large quantities.

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  • Method for quickly extracting genome DNA of riemerella anatipestifer, and kit
  • Method for quickly extracting genome DNA of riemerella anatipestifer, and kit

Examples

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Embodiment 1

[0030] A method for rapidly extracting Riemerella anatipestifer genomic DNA, the specific steps are as follows:

[0031] (1) Collection of bacterial cells: Take 1.5 mL overnight culture liquid of Riemerella anatipestifer RA1 strain in the logarithmic growth phase, add it to a 1.5 mL centrifuge tube, centrifuge at room temperature 12000 r / min for 3 min, discard Supernatant, collect bacteria, if you need to increase the concentration of bacterial genomic DNA, you can repeat the sampling;

[0032] (2) Cell lysis: Add 200 μL of lysis buffer (40 mM Tris-acetate pH 7.8, 20 mM sodium-acetate, 1 mM EDTA, 1% SDS)) to the pellet, gently pipette several times, and fully resuspend For bacteria, let the centrifuge tube stand at room temperature for 2 minutes to fully lyse the bacteria;

[0033] (3) Precipitate bacterial protein: add 70 μL of 5 M NaCl solution to the resuspension, mix thoroughly, and centrifuge at 12,000 r / min at 4°C for 10 min; the addition of 5 M NaCl is helpful for the ...

Embodiment 2

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Abstract

The invention discloses a method for quickly extracting a genome DNA (Deoxyribo-Nucleic Acid) of riemerella anatipestifer, and a kit. The method comprises the following steps: 1) lysing bacteria; 2) precipitating bacterial proteins; 3) removing the bacterial proteins; 4) precipitating the genome DNA; 5) deionizing; and 6) dissolving the DNA, wherein a bacterial lysis buffer solution used in the step 1) contains 38-42 mM of Tris-acetate having the pH of 7.8, 18-22 mM of sodium-acetate, 0.8-1.2 mM of EDTA (Ethylene Diamine Tetraacetic Acid) and 0.8-1.2% of SDS (Sodium Dodecyl Sulfonate). The extraction method provided by the invention is high in extraction efficiency, and the genome DNA of riemerella anatipestifer extracted by the method is high in purity, which lays a strong foundation for subsequent molecular biologicalstudy; and the extraction method is simple and feasible to operate, and high in practicability.

Description

technical field [0001] The invention relates to the field of molecular biology of microorganisms, and in particular provides a method for efficiently extracting high-molecular-weight genomes from the genome of Merella bacterium. The genome extracted by this method can meet the needs of many molecular biology analysis. Background technique [0002] Riemerella anatipestifer (RA) is a Gram-negative bacterium that is non-motile and does not form spores. It mainly causes infectious sepsis in domestic ducks, turkeys and many other birds. Since Guo Yupu and others confirmed the existence of the disease in my country in 1982, Riemerella anatipestifer has been a major disease that has endangered the duck industry in my country. The morbidity and mortality of this disease are all very high, and its incidence rate can reach more than 90%, and the mortality rate can reach 75%. Stunting. The disease is widely prevalent in the world and is currently one of the most important and seriou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12R1/01
Inventor 袁建丰李林林孙敏华董嘉文向蓉胡奇林
Owner INST OF ANIMAL HEALTH GUANGDONG ACADEMY OF AGRI SCI
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