Soil DNA extraction method for analyzing microbial community structure of polycyclic aromatic hydrocarbon polluted land

A microbial community, polycyclic aromatic hydrocarbon technology, applied in the field of soil microbial molecular ecology, to achieve the effect of strong applicability, high safety and simple extraction method

Inactive Publication Date: 2017-10-17
BEIJING MUNICIPAL RES INST OF ENVIRONMENT PROTECTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The reported methods for extracting total DNA of microbial genomes from different environmental samples have certain limitations.

Method used

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  • Soil DNA extraction method for analyzing microbial community structure of polycyclic aromatic hydrocarbon polluted land
  • Soil DNA extraction method for analyzing microbial community structure of polycyclic aromatic hydrocarbon polluted land
  • Soil DNA extraction method for analyzing microbial community structure of polycyclic aromatic hydrocarbon polluted land

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Embodiment 1

[0048] The content of the present invention will be further explained and illustrated below in conjunction with specific examples.

[0049] The test soil for the method of this embodiment is the soil on the polluted site of a comprehensive coking plant production enterprise. This enterprise uses coal as the main raw material and has produced gas, coke and coal chemical products since the 1950s. The incomplete combustion of fossil fuels in the production workshop and the processing of chemical products such as gas and coke may lead to the emission of polycyclic aromatic hydrocarbons. Yu Nian is a typical PAH-contaminated soil.

[0050] The reagent preparation method used

[0051] (1) β-mercaptoethanol-soil pretreatment buffer:

[0052] 50mM Tris, 20mM EDTA, 100mM NaCl, 10mg / mL PVP, pH10;

[0053] After preparation, add β-mercaptoethanol with a volume ratio of 1.5%, and mix well;

[0054] (2) 1×PBS buffer:

[0055] 8.00g NaCl, 0.20g KCl, 1.44g NaCl 2 HPO 4 , 0.24g KH 2 P...

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Abstract

The invention discloses a soil DNA extraction method for analyzing the microbial community structure of a polycyclic aromatic hydrocarbon polluted land. The method comprises the following specific steps: 1, preparation of a reagent; 2, extraction of a general DNA. The extraction method is simple, convenient to operate, high in applicability and high in safety. For the characteristics of severe polycyclic aromatic hydrocarbon pollution, long-term pollution history, high humus content and the like in soil, firstly, beta-mercaptoethanol with an anti-oxidization effect at a volume ratio of 1.5 percent is added into a soil pretreatment buffer solution to reduce influence of impurities in the soil on DNA extraction; then a relatively mild method for crushing microbial cells through mechanical force of glass beads is adopted, so that the DNA is fully released; furthermore, no high shear force is caused on the DNA, so that the extracted genome DNA is relatively high in intactness.

Description

technical field [0001] The invention belongs to the technical field of soil microbial molecular ecology, and in particular relates to a soil DNA extraction method for analyzing the microbial community structure of a polycyclic aromatic hydrocarbon polluted site. Background technique [0002] In the process of rapid industrialization in our country, due to the lack of people's awareness of environmental protection, the environment on which we live has been severely damaged. Soil, as a natural resource for human beings, has not been spared, and the problem of soil environmental pollution has become more and more acute. Among them, the pollution phenomenon of polycyclic aromatic hydrocarbons (Polycyclic aromatic hydrocarbons, polycyclic aromatic hydrocarbons) with strong carcinogenic, teratogenic and mutagenic effects is very common. Coal chemical companies and coking companies that use coal as the main raw material for production activities are one of the important factors tha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 张丹姜林夏天翔
Owner BEIJING MUNICIPAL RES INST OF ENVIRONMENT PROTECTION
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