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Specific primers for quantitative detection of red-eared slider biomass in environment and detection method and application of specific primers

A quantitative detection, red-eared turtle technology, applied in biochemical equipment and methods, microbial 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: 2019-12-27
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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  • Specific primers for quantitative detection of red-eared slider biomass in environment and detection method and application of specific primers
  • Specific primers for quantitative detection of red-eared slider biomass in environment and detection method and application of specific primers
  • Specific primers for quantitative detection of red-eared slider biomass in environment and detection method and application of specific primers

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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 COI gene sequence (NCBI sequence number: KM216748.1) in the mitochondrial DNA of the red-eared turtle from the NCBI website, and use the COI gene sequence as a template to design specific PCR primers for detecting the red-eared turtle. This example obtained a total of Three pairs of 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]

[0034] 2.2 Validation of primers

[0035] In order to verify the effectiveness of the above primers, in the present embodiment, the red-eared turtle and other 6 kinds of com...

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Abstract

The invention relates to specific primers for quantitative detection of the red-eared slider biomass in an environment and a detection method and application of the specific primers. A red-eared slider mitochondrial COI gene is selected as a target gene, the specific amplification primers KM216748F2 and KM216748R2 are developed and obtained, the specific primers are used for being combined with addPCR technology, and the quantitative detection of the red-eared turtle biomass in the environment is realized. Compared with the prior art, the primers can be combined with the ddPCR technology, specific amplification of the red-eared slider mitochondrial gene is realized, the specific primers have the characteristic of high pertinence, in addition, the specific primers do not depend on a standard curve in the detection process, the interference of inhibitors in background DNA and the influence of PCR amplification efficiency changes in different samples and during tests can be avoided, andhigher sensitivity and accuracy are achieved.

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 mainland of my country via Hong Kong 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 artifi...

Claims

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Application Information

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Patent Type & Authority Applications(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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