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A real-time fluorescent quantitative PCR detection method and primers used for the cu/zn-sod gene of Snakehead sinensis

A real-time fluorescence quantification technology, which is applied in the field of fluorescence quantitative PCR detection, can solve the problems that the detection method has not been systematically reported, and achieve the effect of high accuracy, simple method and high reliability

Active Publication Date: 2021-09-17
ZHEJIANG OCEAN UNIV
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Problems solved by technology

[0004] However, there is no systematic report on the real-time fluorescent quantitative PCR detection method of Cu / Zn-SOD gene in Channa sinensis.

Method used

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  • A real-time fluorescent quantitative PCR detection method and primers used for the cu/zn-sod gene of Snakehead sinensis

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Effect test

Embodiment

[0038] 1) Design of real-time fluorescent quantitative primers for Cu / Zn-SOD of the order of Channa sinensis:

[0039] Referring to the open reading frame sequence (SEQ ID NO.1) of the Cu / Zn-SOD gene of Channa chinensis, primers with suitable product fragment size and excellent parameters were designed. The open reading frame sequence of Channa chinensis Cu / Zn-SOD gene is shown in SEQ ID NO.1, which contains 465 nucleotide bases, and the nucleotide sequence shown in SEQ ID NO.1 is obtained from the inventor's previous research The Cu / Zn-SOD gene sequence of Channa sinensis. Using the Primer Premier5 software, referring to the open reading frame sequence (SEQ ID NO.1) of the Cu / Zn-SOD gene of Channa sinensis, the product fragment size is designed to be 80-200bp, and the parameters are excellent (no mismatches, no dimers) , no hairpin structure, annealing temperature around 60°C) primer sequence;

[0040] The designed real-time fluorescence quantitative primer sequence of Cu / Z...

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Abstract

The invention relates to the technical field of fluorescent quantitative PCR detection methods. In order to solve the problem that there is no real-time fluorescent quantitative PCR detection method for the Cu / Zn-SOD gene of S. sinensis, the invention provides a Cu / Zn ‑SOD gene real-time fluorescent quantitative PCR detection method and primers used. The method comprises: 1) design of real-time fluorescent quantitative primers for the gene Cu / Zn-SOD of Channa sinensis; 2) design of real-time fluorescent quantitative primers for internal reference gene β-Actin of Channa sinensis; 3) different Reverse transcription of tissue cDNA; 4) real-time fluorescent quantitative PCR detection of Cu / Zn-SOD gene of Channa sinensis. The present invention can realize the real-time fluorescent quantitative PCR detection of the Cu / Zn-SOD gene of S. sinensis; the method is simple, has high accuracy and high reliability; and can realize the detection of different tissues of S. chinensis.

Description

technical field [0001] The invention relates to the technical field of fluorescent quantitative PCR detection methods, in particular to a real-time fluorescent quantitative PCR detection method and primers for the Cu / Zn-SOD gene of Channa sinensis. Background technique [0002] Chinese snakehead (Bostrychus sinensis), belonging to the genus Bostrychus sinensis of the family Perciformes, is a small fish in brackish water and warm water in estuaries. The fish has delicate meat, high nutritional value, and has the effect of accelerating wound healing. It is one of the most valuable edible fish. At present, Snakena sinensis has become one of the important seawater economically farmed fishes in Fujian, Guangdong, Guangxi and other regions of my country. With the continuous deterioration of environmental pollution and aquaculture water bodies, Sinensis sinensis is constantly threatened by pathogens, heavy metals, organic pollutants, etc., resulting in oxidative damage to the body...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6851C12N15/11
CPCC12Q1/6851C12Q2531/113C12Q2563/107C12Q2545/101C12Q2561/113
Inventor 沈斌魏可卢德政巩壮
Owner ZHEJIANG OCEAN UNIV
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