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Extracting reagent, extracting method and kit for cell-bacterial parasite total DNA and application

An extraction method and technology of parasitic bacteria, applied in the field of molecular biotechnology and medicine and health, can solve the problems of unable to meet the needs of extraction efficiency and quality, low total DNA content, etc., achieve good practical application value, and improve the effect of extraction purity

Active Publication Date: 2019-05-31
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been confirmed by experiments that the total DNA content of cells and their parasites extracted from the currently developed experimental method for extracting total nucleic acid DNA of hosts and parasites is low (1.5×10 6 The extraction amount of Drosophila S2 cells and the parasites in them is 200-500ng), which cannot meet the needs of the experiment for extraction efficiency and quality

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] This embodiment provides a reagent for extracting total DNA of cell-parasitic bacteria, the extraction reagent includes proteinase K and lysozyme.

[0033] Lysozyme, also known as muramidase or N-acetylmuramideglycanohydrlase, is an alkaline enzyme that can hydrolyze mucopolysaccharides in pathogenic bacteria. Mainly by destroying the β-1,4 glycosidic bond between N-acetylmuramic acid and N-acetylglucosamine in the cell wall, the insoluble mucopolysaccharides in the cell wall are decomposed into soluble glycopeptides, resulting in the escape of the contents of the cell wall Bacteria dissolve.

[0034] Proteinase K is a powerful proteolytic enzyme isolated from Candida albicans, which has high specific activity and is a key reagent for DNA extraction.

[0035] The above reagent for extracting the total DNA of the cell-parasite can be used in extracting the total DNA of the cell-parasite.

[0036] The above reagent for extracting the total DNA of the cell-parasite can b...

Embodiment 2

[0038] This embodiment provides a reagent for extracting the total DNA of cells-parasites, including proteinase K and lysozyme; the extraction reagent also includes Tris-Cl saturated phenol solution, chloroform, isoamyl alcohol and TE buffer.

[0039] The above reagent for extracting the total DNA of the cell-parasite can be used in extracting the total DNA of the cell-parasite.

[0040] The above reagent for extracting the total DNA of the cell-parasite can be used in the preparation of the kit for extracting the total DNA of the cell-parasite.

Embodiment 3

[0042] This embodiment provides a kit for extracting total DNA from cells-parasites, and the extraction reagents include proteinase K and lysozyme.

[0043] Of course, the kit can also include a buffer reagent for extracting the total DNA of the cell-parasite, and the buffer reagent is prepared from at least one of Tris-Cl saturated phenol solution, chloroform, ethanol, isoamyl alcohol and TE solution.

[0044] The above-mentioned extraction kit of cell-parasite total DNA can be applied in extracting cell-parasite total DNA.

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PUM

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Abstract

The invention provides an extracting reagent, extracting method and kit for cell-bacterial parasite total DNA and application, and belongs to the field of molecular biotechnology and medicine and hygiene. According to the extracting reagent, extracting method and kit for the cell-bacterial parasite total DNA and the application, by fusing lysozyme and proteinase K, an extracting buffering solutionis prepared, the kit is prepared accordingly, the efficiency of extracting the cell-bacterial parasite total DNA can be well improved, and the extracting purity can be well improved; the extracting reagent, extracting method and kit for the cell-bacterial parasite total DNA and the application have high practical application value for medicine and food hygiene.

Description

technical field [0001] The invention relates to the fields of molecular biology technology and medicine and hygiene, in particular to a reagent, kit, application and extraction method for total DNA of cell-parasitic bacteria. Background technique [0002] At present, total DNA extraction kits are mainly aimed at the extraction of genomic DNA, mitochondrial DNA, or viral DNA in tissues, blood, and animal secretions. There are very few kits specifically for the extraction of total DNA from host cells and pathogenic bacteria. The efficiency is low. In the study of the interaction between the host and the parasite, it is particularly important to effectively obtain the DNA or RNA of the cell and the parasite as a whole. It has been confirmed by experiments that the total DNA content of cells and their parasites extracted from the currently developed experimental method for extracting total nucleic acid DNA of hosts and parasites is low (1.5×10 6 The extraction amount of Drosop...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 田铃吴坤钟曹阳黄志君
Owner SOUTH CHINA AGRI UNIV