Analysis method for diversity of microbes and abundance of species in coal seam water

An analytical method and microbial technology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of low sensitivity, cumbersome operation, time-consuming and laborious, etc., and achieve the effect of high sensitivity and simple operation

Inactive Publication Date: 2014-08-13
SHANXI JINCHENG ANTHRACITE COAL MINING GRP CO LTD
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Problems solved by technology

[0006] In order to solve the limitations of existing research on microbial diversity in the environment, the present invention requires a large amount of samples, cumbersome operation, time-consuming and laborious, low sensitivity, and cannot effectively detect species with

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  • Analysis method for diversity of microbes and abundance of species in coal seam water
  • Analysis method for diversity of microbes and abundance of species in coal seam water
  • Analysis method for diversity of microbes and abundance of species in coal seam water

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Embodiment 1

[0018] Embodiment 1: a kind of analysis method of microbial diversity and species abundance in coal seam water, concrete steps are as follows:

[0019] (1) Coal seam water sample collection: Collect coal seam water samples under anaerobic conditions. The device used to store water samples is sterilized and placed on an ultra-clean workbench, and the other end of the catheter connected to the nitrogen cylinder or argon cylinder is connected to filter After the device is placed in the device, the valve of the gas cylinder is opened, the volume of the gas introduced is 5-10 times larger than the volume of the device, and the air in it is replaced; then cysteine ​​with a final concentration of 0.05% is added as a reducing agent, -20 o C airtight storage;

[0020] (2) Enrichment of microbial cells and DNA extraction: In order to ensure sufficient DNA extraction, the 3L water sample collected on the aseptic operating table was filtered through a vacuum filtration device, and collect...

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Abstract

The invention belongs to the field of research on microbes in stratum water and provides an analysis method for diversity of microbes and abundance of species in coal seam water to overcome the problems that research on diversity of microbes in an environment has great limitation, species with low abundance in a sample cannot be effectively detected, species with high similarity in the sample are difficult to distinguish, only a minority of microbe species in the sample can be revealed, etc. The method comprises the following steps: acquisition of a coal seam water sample; enrichment of microbial thalli in the water sample and extraction of total DNA of the microbial thalli; amplification of the hypervariable region of 16SrDNA with specific primers; sequencing analysis with an Illumina sequencing platform; and processing of original data of sequencing so as to obtain diversity and abundance of microbes in the water sample. The method can effectively detect species with low abundance in the sample, identifies more than 400 genuses of bacteria in coal seam water, has obvious superiority in research on species and abundance of microbes in coal seam water and is applicable to analysis of diversity of microbes and abundance of species in environments like coal seams and oil gas or shale gas strata.

Description

technical field [0001] The invention belongs to the research field of formation water microorganisms, in particular to a method for analyzing the diversity and species abundance of microorganisms in coal seam water. Background technique [0002] Coalbed gas, commonly known as gas, is a self-storage natural gas generated during coal formation and stored in coal seams and surrounding rocks in an adsorbed and free state, in which the volume fraction of CH4 is up to 95%. According to the 2006 national oil and gas resource evaluation, the coalbed methane resources with a buried depth of less than 2000 m in my country reached 36.81×1012 m 3 , which is basically equivalent to the prospective resources of natural gas in my country. With the rapid growth of the national economy, my country's demand for energy is increasing. As unconventional natural gas, coalbed methane is an important supplementary source of natural gas resources in my country. The development and utilization of ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04
CPCC12Q1/6869
Inventor 王保玉刘建民白建平韩作颖刘健苗彪赵娜
Owner SHANXI JINCHENG ANTHRACITE COAL MINING GRP CO LTD
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