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Method for analyzing diversity of biological enzyme genes in compost

An analysis method and bio-enzyme technology, applied in biochemical equipment and methods, microbial measurement/inspection, analysis materials, etc., can solve the problems of increasing experimental costs, increasing workload, and not suitable for the separation and diversity of multiple functional genes Research and other issues to achieve the effect of saving experimental costs, improving work efficiency, and low cost

Inactive Publication Date: 2009-10-07
HUNAN UNIV
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Problems solved by technology

[0004] At present, the research on the separation and diversity of functional genes in the environment should be combined with the method of constructing a clone library, but this method is not suitable for the research on the separation and diversity of various functional genes in the composting process. The reasons are: construction A clone library must pick dozens or even hundreds of clones at a time, and composting is a dynamic process that is constantly changing. Therefore, if you want to isolate multiple functional genes in the composting process and conduct subsequent diversity research It is necessary to construct multiple clone libraries, which not only greatly increases the workload, but also increases the experimental cost to a certain extent

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  • Method for analyzing diversity of biological enzyme genes in compost
  • Method for analyzing diversity of biological enzyme genes in compost
  • Method for analyzing diversity of biological enzyme genes in compost

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Embodiment

[0049] The compost sample to be analyzed by the analysis method of this embodiment is taken from a 20L experimental-scale composting device. The compost materials mainly include 1.074kg of rice straw, 0.9kg of vegetable leaves, 0.216kg of leaves, 0.24kg of bran and 0.721kg of soil. 3 cm below the compost surface, three samples were taken in total (numbered 1, 2, 3 respectively), each sample was 1 g, and the sampling time was 12 days after composting treatment (for compost maturity period samples), the temperature was 38 °C, and the pH value was 8.32, the water content is 58%, and the organic matter content is 20.3%. Among them, the pH value is measured by the glass electrode method, the moisture content is measured by repeated baking at 105°C until constant weight is measured, and the organic matter content is measured by the potassium dichromate method.

[0050] Adopt analysis method of the present invention to carry out the analysis of laccase gene diversity in compost to ab...

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Abstract

The invention discloses a method for analyzing diversity of biological enzyme genes in a compost, comprising the following steps of: at first, extracting microbial total DNA in the compost, wherein, the extraction procedure also comprises three steps of dehumification of a sample, crude extraction of the DNA and purification of the DNA; afterwards, utilizing a PCR amplification program to amplify specific biological enzyme genes; then implementing denaturing gel gradient electrophoresis for 12-14 hours in 1XTAE buffer solution at the constant temperature of 60 DEG C and under the constant voltage of 100V; and finally, analyzing a denaturing gel gradient electrophoresis map to determine the diversity of the specific biological enzyme genes in the compost. The inventive analysis method has the advantages of high flux, low cost, great simplicity and being visual, in order to better represent microbial population having a function degradation in the compost.

Description

technical field [0001] The invention relates to a method for gene diversity, in particular to an analysis method for gene diversity in the total DNA of composting microorganisms. Background technique [0002] Composting is a system in which microorganisms are used to decompose biodegradable solid waste to achieve its final treatment. Therefore, a full understanding of the microbiological principles can provide a basis for improving the treatment process and improving treatment efficiency. The traditional methods for microbial community research on compost are mainly microbial culture and pure species isolation techniques. Because the vast majority of microorganisms in the environment cannot be studied by culture methods, and there are many kinds of microorganisms in compost, which are often in a state of dynamic change, and culture research cannot reflect the dynamic changes of microbial populations in the system. Therefore, such research methods cannot meet the needs of com...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N27/447G01N21/78
Inventor 范长征曾光明杨春平梁婕杨朝晖李凤李贞肖勇
Owner HUNAN UNIV
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