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
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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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