Biomembrane-alpha proteobacteria quantitative determination method

A quantitative detection method, the technology of proteobacteria, applied in the field of molecular biology, can solve the problems of time-consuming microbial detection technology, difficulty in meeting the dynamic monitoring of microbial community structure changes, and difficulty in determining microbial diversity.

Inactive Publication Date: 2014-11-19
上海浦东威立雅自来水有限公司 +1
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Problems solved by technology

Furthermore, the culture-based microbial detection technology cannot meet the needs of analyzing the structure-function relationship of microbial communities due to its high selectivity, time-consuming, and labor-intensive shortcomings.
In addition, many organisms are not cultivable, and it is difficult to determine the microbial diversity in the ecosystem; traditional separation and counting techniques are also difficult to meet the requirements of dynamic monitoring of microbial community structure changes

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  • Biomembrane-alpha proteobacteria quantitative determination method
  • Biomembrane-alpha proteobacteria quantitative determination method

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Embodiment Construction

[0077] In order to make the measures, creative features, goals and effects achieved by the technology of the present invention easy to understand, the present invention will be further described below in conjunction with specific illustrations.

[0078] A quantitative detection method for biofilm α-proteobacteria, comprising the steps of:

[0079] Step 1, sample pretreatment;

[0080] Firstly, the standard sample of α-proteobacteria is pretreated, that is, the recovery operation of the standard sample of α-proteobacteria is carried out to obtain a freshly activated standard sample liquid. The specific operation is as follows;

[0081] Take out the α-proteobacteria standard sample stored at -80°C, dissolve it to room temperature, and mix well;

[0082] Further, take 20 μl of the mixed liquid α-proteobacteria standard sample and add it to 2ml LB liquid medium sterilized at 121°C for 25 minutes before use;

[0083] Further, it was cultured in a constant temperature shaker at 37...

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Abstract

The invention belongs to the technical field of molecular biology, and provides a biomembrane-alpha proteobacteria quantitative determination method. The method comprises the following steps: making a sample of alpha-proteobacteria, recovering the sample of alpha-proteobacteria, preparing Escherichia coli competent cell, conversing the alpha-proteobacteria sample to competence bacteria; culturing by selecting bacterium of the sample of alpha-proteobacteria, extracting the plasmid of the sample of alpha-proteobacteria, determining the quantification double chain DNA content of alpha-proteobacteria, pretreating the biomembrane sample to be measured; and performing real-time fluorescence quantification PCR detection. The method has the advantages of fast analysis speed, less restriction factor, high accuracy, and reduced subjective factor influence, and provides support for microbe stability of a drinking water system as well as scientific basis for purifying water quality of a feed pipe net.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a quantitative detection method for α-proteobacteria in biofilms that can meet the requirements for dynamic monitoring of changes in microbial community structure and provide more accurate, comprehensive and sensitive quantitative detection data. Background technique [0002] Proteobacteria is the largest phylum of bacteria, including many pathogenic bacteria, such as Escherichia coli, Salmonella, Solitary bacteria, Helicobacter, etc. Proteobacteria are Gram-negative bacteria. According to the difference of rRNA, proteobacteria are usually divided into 5 groups, which are named after the Greek letters α, β, γ, δ and ε. Whether as symbionts or pathogens, proteobacteria closely interact with eukaryotes. In urban water supply systems, proteobacteria are dominant. In addition to photosynthetic species, α-proteobacteria also have species that metabolize C1 compo...

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/06
CPCC12Q1/6851C12Q2545/114C12Q2561/113C12Q2563/107
Inventor 戴婕王雪峰张丽丽李伟英张东王铮
Owner 上海浦东威立雅自来水有限公司
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