Molecular detection method of Alicyclobacillus acidoterrestris

A Bacillus and molecular detection technology, applied in the field of microbial detection, can solve the problems of high cost, consumption, complicated steps, etc., and achieve the effect of avoiding pollution

Pending Publication Date: 2020-09-15
HEBEI AGRICULTURAL UNIV.
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Problems solved by technology

Including probe design and the process of using the probe to generate circular DNA, making the RCA reaction take longer, more complicated steps, and more expensive

Method used

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  • Molecular detection method of Alicyclobacillus acidoterrestris
  • Molecular detection method of Alicyclobacillus acidoterrestris
  • Molecular detection method of Alicyclobacillus acidoterrestris

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Embodiment Construction

[0036] The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only some, not all, embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

[0037] A molecular detection method for Alicyclobacillus acidic soil, under the action of Bca DNA polymerase and RNase H enzyme, at a temperature of 60-65°C, the amplification reaction of SPIA only needs one mixed primer to complete, Specifically include the following steps:

[0038] Such as figure 1 At the beginning of the reaction, the mixed primer and the chain-terminating polynucleotide (blocker) are respectively combined with the corresponding positions ...

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Abstract

The invention discloses a molecular detection method of Alicyclobacillus acidoterrestris. Under the action of Bca DNA polymerase and RNase H enzyme, the amplification reaction of SPIA can be completedonly by one mixed primer at the temperature of 60-65 DEG C. According to the invention, the dependence of the RCA method on cyclized DNA and the false positive phenomenon which is extremely easy to occur in LAMP can be solved. Therefore, the SPIA has higher detection efficiency for the RCA, and has better stability for the LAMP. As a novel nucleic acid isothermal amplification technology, the molecular detection method is more perfect, and the advantages of the isothermal amplification technology are more prominent.

Description

technical field [0001] The invention relates to the technical field of microbial detection, in particular to a molecular detection method for Alicyclobacillus acidic soil. Background technique [0002] Nucleic acid in vitro amplification technology is the basis of molecular biology research and an important method of biological analysis. Through nucleic acid amplification, the target gene can be amplified and separated, so that it can be used in clinical medical diagnosis, gene mutation monitoring, molecular diagnosis, and food safety detection. And environmental safety monitoring and other fields have important uses. [0003] With the development of molecular biology and the wide application of biophysics in this field, nucleic acid amplification technology has received more and more attention. Common nucleic acid amplification techniques include polymerase chain reaction (PCR), loop-mediated isothermal amplification (LAMP) and rolling circle isothermal amplification (RCA)...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6844C12Q1/689C12R1/01
CPCC12Q1/6844C12Q1/689C12Q2521/327C12Q2531/119C12Q2547/10C12Q2563/107
Inventor 马晓燕张伟张蕴哲张先舟苑宁杨粤
Owner HEBEI AGRICULTURAL UNIV.
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