Extraction method of mycobacterium tuberculosis DNA

A technology of Mycobacterium tuberculosis and extraction method, which is applied in the direction of DNA preparation, recombinant DNA technology, and microbial-based methods, and can solve problems such as unstable extraction effect, poor precipitation effect, and poor dispersion degree

Active Publication Date: 2019-08-27
SHENZHEN UNI MEDICA TECH
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Problems solved by technology

[0005] The purpose of the present invention is to provide a method for extracting Mycobacterium tuberculosis DNA, aiming to solve the problem of poor dispersion of cells with high aggregation degree, unstable extraction effect and low precipitation efficiency in the existing Mycobacterium tuberculosis DNA extraction method, especially It is a technical problem such as poor precipitation effect on a small amount of samples

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  • Extraction method of mycobacterium tuberculosis DNA

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Embodiment 1

[0072] A method for extracting Mycobacterium tuberculosis DNA, comprising the following steps:

[0073] (1) Scrape Mycobacterium tuberculosis colonies from L-J (Loewenstein-Jensen) solid medium, put them into TE buffer solution with 500ul, 80°C water bath, 30 minutes to kill bacteria, and get inactivated Mycobacterium tuberculosis.

[0074] (2) For the inactivated Mycobacterium tuberculosis, perform vortex oscillation dispersion treatment: ① Ensure that the total amount of inactivated strains and 1×TE in the 1.5ml EP tube is 500ul, if insufficient, add 1×TE to make up . ②Add 3-4 glass beads to the EP tube, and vortex fully for 2-3 minutes to disperse the bacteria. ③ According to the amount of bacteria in the EP tube after vortexing, 1×TE can be added appropriately for dilution to obtain the vortex dispersion product.

[0075] (3) Ultrasonic dispersion of the vortex dispersion product: ①Pipe and mix the bacterial solution in the EP tube to prevent bacterial strain precipitat...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to an extraction method of mycobacterium tuberculosis DNA, comprising the following steps: obtaining mycobacterium tuberculosis, and inactivating the mycobacterium tuberculosis to obtain the inactivated mycobacterium tuberculosis; firstly performing vortex oscillation dispersion treatment on the sterilized mycobacterium tuberculosis is subjected to, and then ultrasonically dispersing to obtain mycobacterium tuberculosis dispersion liquid; performing wall breaking treatment on the mycobacterium tuberculosis dispersion liquid to obtain a wall breaking product; performing first extraction on the wall breaking product by a CTAB/NaCl solution, and then performing second extraction by using a mixed liquor of phenol, chloroform and isoamyl alcohol to obtain an extracted product; precipitating the extracted product by isopropanol and glycogen to obtain mycobacterium tuberculosis DNA. The extraction method disclosed by the invention has the advantages of high extraction quality and easiness in operation, and is suitable for being carried out in an ordinary biological laboratory.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for extracting DNA of mycobacterium tuberculosis. Background technique [0002] Mycobacterium tuberculosis (Mycobacterium tuberculosis) is the pathogenic bacterium that causes tuberculosis, which can invade various organs of the whole body, and pulmonary tuberculosis is the most common. Tuberculosis is still an important infectious disease so far. Therefore, the timely, efficient and specific detection of Mycobacterium tuberculosis has become an effective means to prevent and treat diseases caused by Mycobacterium tuberculosis. Although some achievements have been made in the biological characteristics, pathogenic mechanism, drug resistance mechanism, diagnostic methods, and development of anti-tuberculosis drugs of Mycobacterium tuberculosis, tuberculosis is still the most prevalent disease that affects human health. As one of the infectious diseases with the highest rate,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6806C12N15/10C12R1/32
CPCC12Q1/6806C12N15/1003Y02A50/30
Inventor 雷湘华郭永超陈莹莹黄歆黄萍
Owner SHENZHEN UNI MEDICA TECH
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