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Kit for extracting gram-positive bacterium genome

A Gram-positive bacteria and genome technology, applied in the biological field, can solve the problems of time-consuming and DNA sample extraction, and achieve the effect of simplifying experimental steps and saving extraction time

Inactive Publication Date: 2015-12-16
FUJIAN NORMAL UNIV
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AI Technical Summary

Problems solved by technology

[0004] Therefore, it is necessary to develop a convenient, fast and practical method suitable for the extraction of gram-positive bacterial genomic DNA, and solve the problem of long extraction time of DNA samples obtained by conventional methods

Method used

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  • Kit for extracting gram-positive bacterium genome

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Embodiment Construction

[0025] figure 1 In, lane 1 is the test result of extracting staphylococcal genomic DNA using this method, lane 2 is the test result of extracting streptococcal genomic DNA using this method, and lane 3 is the test result of extracting actinomycete genomic DNA using this method . Lane 4 is the DL4500marker from TAKARA. It can be seen from the figure that the present invention can quickly extract genomic DNA of Gram-positive bacteria.

[0026] A kit for extracting gram-positive bacteria genome, the specific steps are as follows:

[0027] (1) Disruption of Gram-positive bacteria cells: Take 3ml of Staphylococcus, Streptococcus, and Actinomycetes overnight culture respectively and add them to 1.5ml centrifuge tubes 3 times, set 3 tubes to repeat, centrifuge at 12000rpm / min for 1min each time Then discard the supernatant. Add 0.25ml ddH to the bacterial pellet 2 O suspend the precipitate, and add 75μl of Buffer A solution, and keep it at 40°C for 30min. Add 50μl Buffer B, 5μl Buffer...

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Abstract

The invention provides a kit for extracting a gram-positive bacterium genome, and belongs to the field of biotechnology. The kit comprises the following specific steps: breaking gram-positive bacterium cells, dissociating DNA and removing impurity protein, adsorbing the DNA by virtue of a DNA adsorption column and removing impurities, and eluting the DNA. Different from a conventional kit for extracting a gram-positive bacterium genome which is relatively complex in extraction step and relatively long in extraction time consumed which generally requires about 12h, the kit for extracting the gram-positive bacterium genome, compared with a conventional kit in method for extracting gram-positive bacterium genome DNA, is capable of extracting the gram-positive bacterium genome DNA for only about 2h, so as to save a lot of extraction time; and meanwhile, the kit can simplify experimental steps, and can be used for extracting the gram-positive bacterium genome DNA more effectively, rapidly and economically.

Description

Technical field [0001] The invention relates to a kit for extracting gram-positive bacteria genome, belonging to the field of biotechnology. technical background [0002] Gram-positive bacteria have thicker cell walls, about 20 to 80 nm. Peptidoglycan is rich in content, with 15-50 layers, each with a thickness of 1nm, accounting for about 50-80% of the dry cell weight. In addition, there are a large number of special components teichoic acid. Teichoic acid is a polymer composed of ribitol or glycerol residues connected to each other via phosphate double bonds. Teichoic acid split wall teichoic acid and membrane teichoic acid. The former is linked to the n-acetyl teichoic acid of peptidoglycan in the cell wall. Membrane teichoic acid, also known as lipoteichoic acid, is linked to the cell membrane, and both ends are free Outside the cell wall. Teichoic acid has strong antigenicity and is an important surface antigen of gram-positive bacteria; it plays a role in regulating the...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 王清水余彦
Owner FUJIAN NORMAL UNIV
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