Specific multiple PCR (polymerase chain reaction) detection method and kit for toxigenic microcystis and kit therefor

A detection kit and detection method technology are applied in the field of toxin-producing Microcystis-specific multiplex PCR detection methods and kits, which can solve the problems of complex detection methods and high instrument requirements for toxin-producing Microcystis, and achieve easy to master, The effect of improving detection accuracy and easy operation

Inactive Publication Date: 2013-03-06
SHANGHAI OCEAN UNIV
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Problems solved by technology

[0004] The technical problem to be solved by the present invention is to provide a mult

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  • Specific multiple PCR (polymerase chain reaction) detection method and kit for toxigenic microcystis and kit therefor
  • Specific multiple PCR (polymerase chain reaction) detection method and kit for toxigenic microcystis and kit therefor
  • Specific multiple PCR (polymerase chain reaction) detection method and kit for toxigenic microcystis and kit therefor

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[0049] In order to make the technical means, creative features, goals and effects achieved by the present invention easy to understand, the present invention will be further described below in conjunction with specific illustrations.

[0050] The following will be described in the form of examples. If there is anything that is not exhaustive, please refer to commonly used experimental manuals, such as the "Molecular Cloning Experimental Manual" and the manufacturer's instructions for the reagents and instruments used. Among them, all chemical reagents are of analytical grade, and the experimental water is filtered by Milli-XQ. All reagents and materials can be obtained from commercial channels. Specifically: agarose was purchased from Shanghai Sangong Biological Company, and the CTAB extraction reagent was hexadecane Trimethylammonium bromide (CTAB), sarkosyl, EDTA, NaCl, Tris-HCl, β-mercaptoethanol (β-mercaptoethano), phenol: chloroform: isoamyl alcohol (PCI, 24:24 : 1) Reage...

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Abstract

Aiming at overcoming the technical problems of complex toxigenic microcystis detection method, high instrument requirement and the like in the prior art, the invention provides a specific multiple PCR (polymerase chain reaction) detection method and kit for toxigenic microcystis. The specific multiple PCR detection method for the toxigenic microcystis comprises the following steps of: (1) sampling microcystis from a detected site, and extracting DNA (deoxyribonucleic acid) of genome; (2) designing and synthesizing the microcystis which comprises seven pairs of primers, i.e. toxigenic and non-toxigenic strain ropC1 gene, toxigenic microcystis mcyA, mcyB, mcyD, mcyE, mcyI and mcyJ gene sections; and (3) respectively carrying out PCR amplification reaction by the designed specific primers, and carrying out electrophoresis gel detection. According to the invention, a plurality of genes are taken as molecular indexes, so that the detection precision can be improved, the method is simple and convenient to operate, and easy to grasp, the microcystis in water body can be quickly detected, and the toxigenic strain can be effectively identified.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a specific multiplex PCR detection method for toxin-producing Microcystis and a kit thereof. Background technique [0002] Since the 1960s, cyanobacteria blooms have been found in Taihu Lake. So far, not only large freshwater lakes such as Dianchi Lake in Yunnan, Taihu Lake in Jiangsu, Chaohu Lake in Anhui, East Lake in Wuhan, and Dianshan Lake in Shanghai have been seriously polluted by cyanobacteria blooms, but many lakes and reservoirs in the middle and lower reaches of the Yangtze River, Yellow River, and Pearl River have also been polluted one after another. Cyanobacterial blooms of varying degrees occurred. Cyanobacteria blooms continue to occur in a large area, which greatly endangers the structure and function of the water ecosystem. The algal toxins produced by some species of algae blooms also pose a direct or potential impact on human health. For example, M...

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04
Inventor 唐晨董丽饶涛何培民贾睿
Owner SHANGHAI OCEAN UNIV
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