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Primer for detecting bostrychus sinensis Mn-SOD gene and real-time fluorescent quantitative PCR method

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

Active Publication Date: 2019-03-05
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 the Mn-SOD gene of Channa sinensis at present

Method used

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  • Primer for detecting bostrychus sinensis Mn-SOD gene and real-time fluorescent quantitative PCR method

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Embodiment

[0038] 1) Design of real-time fluorescent quantitative primers for the gene Mn-SOD of the order Sinensis chinensis

[0039] Referring to the open reading frame sequence (SEQ ID NO.1) of the Mn-SOD gene of Channa chinensis, the primers with suitable product fragment size and excellent parameters were designed. The open reading frame sequence of Mn-SOD gene of Channa sinensis is shown in SEQ ID NO.1, which contains 678 nucleotide bases. The nucleotide sequence shown in SEQ ID NO.1 comes from the Chinese Sequence of the Mn-SOD gene of Channa sinensis. Utilizing the Primer Premier5 software, referring to the open reading frame sequence (SEQ ID NO.1) of the Mn-SOD gene of S. sinensis, the product fragment size was designed to be 150-300bp, and the parameters were excellent (no mismatch, no dimer, no Hairpin structure, annealing temperature around 60°C) primer sequence.

[0040] The sequence of the designed real-time fluorescent quantitative primer pair for Mn-SOD gene of Channa s...

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Abstract

The invention relates to the technical field of a fluorescent quantitative PCR detection method, in particular to a primer for detecting a bostrychus sinensis Mn-SOD gene and a real-time fluorescent quantitative PCR method. According to the method, the real-time fluorescent quantitative PCR technology is used for analyzing an Mn-SOD gene expression amount of different tissues of the bostrychus sinensis, and GAPDH is used as an internal reference gene to design an internal reference gene primer and a target gene primer; and the reverse transcription cDNA is used as a template to detect an expression level of the target gene on a ABI 7500 fast real-time PCR system, the detection result is statistically analyzed, so that the expression and distribution situation of the Mn-SOD gene in different tissues of the bostrychus sinensis can be rapidly and accurately detected, and the scientific evidence is provided for using and researching the gene.

Description

technical field [0001] The invention relates to the technical field of fluorescent quantitative PCR detection methods, in particular to a primer and a real-time fluorescent quantitative PCR method for detecting the Mn-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 c...

Claims

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Application Information

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