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Rapid molecule detecting method for microflora composition in waste water biological treatment reactor

A microbial community and biological treatment technology, applied in the direction of microbial determination/inspection, biological water/sewage treatment, biochemical equipment and methods, etc. effect of change

Inactive Publication Date: 2009-07-08
NANJING UNIV
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Problems solved by technology

At present, there are patent applications for using DGGE technology to analyze the microbial community in urban sewage plant sludge and industrial wastewater treatment, such as the method for analyzing the microbial community composition of urban sewage plant sludge proposed by Ji University (CN1584051), and the industrial wastewater invented by Shanghai Jiaotong University Functional flora-specific molecular detection method in treatment (CN1995389), but does not involve how to further shorten the detection time using DGGE technology to detect microbial communities in wastewater biological treatment more quickly

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  • Rapid molecule detecting method for microflora composition in waste water biological treatment reactor
  • Rapid molecule detecting method for microflora composition in waste water biological treatment reactor
  • Rapid molecule detecting method for microflora composition in waste water biological treatment reactor

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Embodiment Construction

[0028] Microbial sample collection, pretreatment and DNA extraction

[0029] Microbial sample collection and pretreatment Sterile bottle collection of five samples 50ml of the above-mentioned device anaerobic reactor UASB upper, middle and lower layer granular sludge, anoxic pool activity, and secondary aerobic pool granular sludge, after collection, store at -20°C freezing. Collect 5 grams of the biological fluidized bed MBBR biofilm sample together with the filler in the conical flask, add 10ml of sterile PBS buffer, shake vigorously to elute the biofilm from the filler until the biofilm is completely eluted. And wash twice with PBS buffer, collect the precipitate. A total of six samples were freeze-dried, poured into a mortar and added liquid nitrogen to grind until powdery, and stored at -20°C.

[0030] Lysozyme treatment and SDS lysis Suspend 50 mg of the precipitate in 1.5 ml of DNA extraction solution (0.1M Tris-HCl, 0.1M Na 2EDTA, 0.1M NaCl, pH8), and 150 μL of lyso...

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Abstract

A rapid molecular detection method of microbial community composition in a wastewater biological treatment reactor includes: collecting the microorganism samples in the wastewater biological treatment reactor for pretreatment, and using the phenol-chloroform method for extracting total genomic DNA; using 16S rDNAV3 area primers to perform PCR amplification to the sample DNA; using denaturing gradient gel electrophoresis to analyzing the PCR amplification products; using software to analyze the obtained DGGE patterns, and identifying the represented microorganism based on the gel cutting sequencing feature DGGE bands; designing and constructing denaturing gradient gel electrophoresis database as the contradistinguish standard of rapid molecular detection based on the microorganism kinds represented by bands, wherein, the denaturing gradient gel electrophoresis integrated database is designed and constructed by gel cutting cloning sequencing the bands after the PCR products is processed after the DGGE separation and then defining the represented microorganism kinds after consanguinity comparison in the Genbank.

Description

technical field [0001] The invention relates to the technology of using PCR-DGGE technology to design and construct DGGE Marker to quickly analyze and detect the composition of microbial communities in samples such as granular sludge, activated sludge, and biofilm in wastewater biological treatment reactors, and belongs to environmental microbial molecular ecology field. Background technique [0002] Wastewater biological treatment technology is currently the main method for treating municipal wastewater and organic industrial wastewater in the world. In order to improve the performance and stability of wastewater treatment, a lot of research has been done on the design and operating conditions of the reactor. The wastewater treatment process mainly relies on the microbial community with activated sludge (including granular sludge) and biofilm as the core to degrade and transform pollutants. However, due to the lack of research techniques and means, people can only use the ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C02F3/34
Inventor 任洪强符波丁丽丽梁睿廖潇逸
Owner NANJING UNIV
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