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Dna Marker for detecting coal geological microbial bacterial species and its preparation method and application

A technology of microorganisms and bacteria, applied in biochemical equipment and methods, measurement/testing of microorganisms, DNA/RNA fragments, etc., can solve the problems of no DNAMarker research reports, etc., to shorten the time-consuming experiments, sharpen the bands, and accelerate The effect of the experimental procedure

Active Publication Date: 2020-11-10
SHANXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, there is no research report on DNAMarker in PCR-DGGE detection of microbial flora in coal geological environment

Method used

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  • Dna Marker for detecting coal geological microbial bacterial species and its preparation method and application
  • Dna Marker for detecting coal geological microbial bacterial species and its preparation method and application
  • Dna Marker for detecting coal geological microbial bacterial species and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] The DNA Marker used to detect coal geological microbial bacterial species, the DNA Marker is composed of 11 DNA fragments a-k, the nucleotide sequence of DNA fragment a is shown in SEQ.ID.NO.1, the nucleotide sequence of DNA fragment b As shown in SEQ.ID.NO.2, the nucleotide sequence of DNA fragment c is as shown in SEQ.ID.NO.3, the nucleotide sequence of DNA fragment d is as shown in SEQ.ID.NO.4, DNA The nucleotide sequence of fragment e is shown in SEQ.ID.NO.5, the nucleotide sequence of DNA fragment f is shown in SEQ.ID.NO.6, and the nucleotide sequence of DNA fragment g is shown in SEQ.ID. Shown in NO.7, the nucleotide sequence of DNA fragment h is shown in SEQ.ID.NO.8, the nucleotide sequence of DNA fragment i is shown in SEQ.ID.NO.9, the nucleotide sequence of DNA fragment j The acid sequence is shown in SEQ.ID.NO.10, and the nucleotide sequence of DNA fragment k is shown in SEQ.ID.NO.11.

[0047] Concrete preparation method comprises the following steps:

[004...

Embodiment 2

[0109] A preparation method for detecting a DNA Marker of coal geological microbial bacterial species, comprising the following steps:

[0110] 1. Extract the genomic DNA in the enrichment medium of the methyl-type methanogenic type and the ethyl-type methanogenic type of the produced water sample, select 2 sets of primers for PCR amplification, and refer to specific implementation scheme 1 for genome extraction and PCR amplification 1.1, 1.2, 1.3 in 1.1, Genomic DNA 0.7% agarose gel electrophoresis verification map is attached figure 1 shown.

[0111] 2. DGGE analysis:

[0112] 2.1 Select the concentration of polyacrylamide gel to be 10%, and the range of gel denaturation is 40% to 60%. Take 18ml each of 40% and 60% gel, add 50μL of TEMED and 40μL of 10% ammonium persulfate respectively, and make gradient mixture The gel can be solidified at room temperature in summer, and it can be solidified in a 37°C incubator in winter. If it is solidified at room temperature, the gel t...

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Abstract

The invention relates to a DNA Marker specially used for coal geology environment bacteria species in molecular detection of denaturing gradient gel electrophoresis (DGGE) for coal geology environment microbial flora, and belongs to the field of microorganism molecular ecology. According to the DNA Marker for detecting the coal geology microorganism bacterium species, the DNA Marker consists of 11 DNA segments a to k, wherein the nucleotide sequence of DNA segment a is as shown in SEQ. ID. No.1; the nucleotide sequence of DNA segment b is as shown in SEQ. ID. No.2; the nucleotide sequence of DNA segment c is as shown in SEQ. ID. No.3; the nucleotide sequence of DNA segment d is as shown in SEQ. ID. No.4; the nucleotide sequence of DNA segment e is as shown in SEQ. ID. No.5; the nucleotide sequence of DNA segment f is as shown in SEQ. ID. No.6; the nucleotide sequence of DNA segment g is as shown in SEQ. ID. No.7; the nucleotide sequence of DNA segment h is as shown in SEQ. ID. No.8; the nucleotide sequence of DNA segment i is as shown in SEQ. ID. No.9; the nucleotide sequence of DNA segment j is as shown in SEQ. ID. No.10; and the nucleotide sequence of DNA segment k is as shown in SEQ. ID. No.11.

Description

technical field [0001] The invention relates to a species DNA Marker specially used for coal geological environment bacteria in denatured gradient gel electrophoresis (DGGE) molecular detection of microbial flora in coal geological environment, belonging to the field of microbial molecular ecology. Background technique [0002] Microorganisms in the coal geological environment refer to the microorganisms detected in coal seam output water samples or coal samples, or the microorganisms detected after enrichment and culture of the above two, which are mainly divided into two categories: bacteria and archaea. They are mainly involved in the process of degrading coal to produce methane. As a clean energy source, coalbed methane has attracted much attention in the world. According to different causes, coalbed methane can be divided into thermogenic gas and biogenic gas. Biogenic gas can be divided into primary biogenic gas and secondary biogenic gas. Primary biogenic gas is form...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/689C12Q1/04C12N15/11C12N15/70
CPCC12N15/70C12Q1/689C12Q2600/166
Inventor 杨秀清陈彦梅王保玉袁宗本吴瑞薇
Owner SHANXI UNIV
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