DNA Marker for detecting coal geology microorganism bacterium species, and preparation method and application

A technology of microorganisms and bacteria, applied in the fields of biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problem of no DNAMarker research report, etc., and achieve shortened experimental time, sharp bands, and accelerated The effect of the experimental process

Active Publication Date: 2017-09-01
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 geology microorganism bacterium species, and preparation method and application
  • DNA Marker for detecting coal geology microorganism bacterium species, and preparation method and application
  • DNA Marker for detecting coal geology microorganism bacterium species, and preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] A DNA Marker for detecting coal geological microbial bacterial species, the DNA Marker consists of 11 DNA fragments a-k, the nucleotide sequence of DNA fragment a is shown in SEQ.ID.NO.1, and the nucleotide sequence of DNA fragment b is shown in SEQ.ID.NO.1 As shown in SEQ.ID.NO.2, the nucleotide sequence of DNA fragment c is shown as SEQ.ID.NO.3, the nucleotide sequence of DNA fragment d is shown as SEQ.ID.NO.4, the 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. As 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, and the nucleotide sequence of DNA fragment j is shown in SEQ.ID.NO.9. 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] The specific p...

Embodiment 2

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

[0110] 1. Extract the methyl-type methane-producing type and ethyl-type methane-producing type of the output water sample to enrich the genomic DNA in the culture medium, select 2 sets of primers for PCR amplification, and refer to specific embodiment 1 for genome extraction and PCR amplification In 1.1, 1.2, 1.3, the verification map of genomic DNA 0.7% agarose gel electrophoresis is as attached figure 1 shown.

[0111] 2. DGGE analysis:

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

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PUM

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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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IPC IPC(8): C12Q1/68C12Q1/04C12N15/11C12N15/70
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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