A primer and real-time fluorescent quantitative PCR method for detecting the mn-sod gene of Channa sinensis

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, high reliability and simple method.

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 the Mn-SOD gene of Channa sinensis at present

Method used

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  • A primer and real-time fluorescent quantitative PCR method for detecting the mn-sod gene of Channa sinensis

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Experimental program
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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 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 Chinese black snakehead (Bostrychus sinensis), which uses real-time fluorescent quantitative PCR technology to analyze Chinese black snakehead The expression of Mn‑SOD gene in different tissues of Channa sinensis, using GAPDH as an internal reference gene, designing internal reference gene primers and target gene primers; using reverse transcribed cDNA as a template, the expression of the target gene on the ABI 7500 Fast Real‑time PCR system It can quickly and accurately detect the expression and distribution of Mn-SOD gene in different tissues of Channa sinensis, and provide a scientific basis for the utilization and research of this 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 Patents(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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