A specific primer for quantitative detection of red-eared turtle biomass in the environment, its detection method and application

A technology for quantitative detection and detection of the environment, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the problems of low sensitivity, accuracy and repeatability, and achieve high sensitivity and accuracy , Accurate quantitative detection of copy number concentration, highly targeted effect

Active Publication Date: 2022-05-03
NANJING INST OF ENVIRONMENTAL SCI MINIST OF ECOLOGY & ENVIRONMENT OF THE PEOPLES REPUBLIC OF CHINA
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Problems solved by technology

[0006] The purpose of the present invention is just to provide a kind of specific primer and its detection method and application of the biomass of red-eared turtle in quantitative detection environment in order to solve the above-mentioned problem, to provide a kind of specific amplification primer sequence, this primer sequence cooperates ddPCR technology, which solves the technical problems of low sensitivity, accuracy and repeatability of existing detection technologies

Method used

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  • A specific primer for quantitative detection of red-eared turtle biomass in the environment, its detection method and application
  • A specific primer for quantitative detection of red-eared turtle biomass in the environment, its detection method and application
  • A specific primer for quantitative detection of red-eared turtle biomass in the environment, its detection method and application

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Embodiment 1

[0027] 1. Materials

[0028] The methods used in this example are conventional methods known to those skilled in the art unless otherwise specified, and the reagents and other materials used are all commercially available products unless otherwise specified.

[0029] 2. Method

[0030] 2.1 Primer design

[0031] Obtain the gene sequence in the mitochondrial DNA of the red-eared turtle from the NCBI website (NCBI sequence number: KM216748.1), and use the gene sequence as a template to design specific PCR primers for the detection of the red-eared turtle. In this example, three pairs were obtained The primers are KM216748-F1 / KM216748-R1, KM216748-F2 / KM216748-R2 and KM216748-F3 / KM216748-R3 respectively. The primer sequences are shown in Table 1 below:

[0032] Table 1: Primer Sequence List

[0033] Primer name sequence KM216748-F1 5'-TCAATTTTCTTCTAGGGTAAATCC-3' KM216748-R1 5'-CAGTAGCCATCATTGAACTAGGAC-3' KM216748-F2 5'-TCTTCTAGGGTAAATCCTATTG-3' ...

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Abstract

The present invention relates to a specific primer for quantitatively detecting the biomass of red-eared turtles in the environment and its detection method and application. The present invention selects the mitochondrial gene of red-eared turtles as the target gene, and develops and obtains a specific amplification primer KM216748F2 and KM216748R2 , and use the specific primers combined with ddPCR technology to achieve quantitative detection of red-eared turtle biomass in the environment. Compared with the prior art, the primer can be combined with ddPCR technology to achieve specific amplification of the mitochondrial gene of the red-eared turtle, which has the characteristics of strong pertinence. In addition, the present invention does not rely on the standard curve during the detection process, and It can avoid the interference of inhibitors in background DNA and the influence of PCR amplification efficiency changes in different samples and test processes, and has higher sensitivity and accuracy.

Description

technical field [0001] The invention belongs to the technical field of environmental molecular detection, and in particular relates to a specific primer for quantitatively detecting the biomass of red-eared turtles in the environment, a detection method and an application thereof. Background technique [0002] The red-eared turtle (Trachemys scripta elegans) is native to the central part of the United States and has been introduced to many countries in Europe, Africa, Asia, and the Americas. It poses a serious threat to the ecological security of the invaded area and is listed as a world species by the International Union for Conservation of Nature (IUCN). One of the top 100 most threatened invasive species. The red-eared turtle was introduced into the inland of China via Hong Kong, my country in the 1980s. At the beginning of the 21st century, this species of artificial breeding has formed a large scale. At the same time, it has entered the wild natural environment due to a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12Q1/6851C12N15/11
CPCC12Q1/6888C12Q1/6851C12Q2531/113C12Q2563/159C12Q2563/107
Inventor 董姗姗刘燕章嫡妮张振华卢晓强于赐刚
Owner NANJING INST OF ENVIRONMENTAL SCI MINIST OF ECOLOGY & ENVIRONMENT OF THE PEOPLES REPUBLIC OF CHINA
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