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Method for interfering real-time fluorescent quantitative PCR reference gene screening of MSTN bovine skeletal muscle satellite cells

A technology of real-time fluorescence quantification and internal reference genes, which is applied in the fields of cell, biology, and tissue engineering, and can solve problems such as patent publications that have not yet been found

Inactive Publication Date: 2021-01-08
TIANJIN AGRICULTURE COLLEGE
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[0004] Through the search, no patent publications related to the patent application of the present invention have been found

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  • Method for interfering real-time fluorescent quantitative PCR reference gene screening of MSTN bovine skeletal muscle satellite cells
  • Method for interfering real-time fluorescent quantitative PCR reference gene screening of MSTN bovine skeletal muscle satellite cells
  • Method for interfering real-time fluorescent quantitative PCR reference gene screening of MSTN bovine skeletal muscle satellite cells

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Embodiment Construction

[0051] Below in conjunction with embodiment, the present invention is further described, and following embodiment is descriptive, not limiting, can not limit protection scope of the present invention with following embodiment.

[0052] The raw materials used in the present invention, if no special instructions, are conventional commercially available products, the methods used in the present invention, if no special instructions, are conventional methods in this area, and the quality of each material used in the present invention is routine use quality.

[0053] A method for interfering with MSTN bovine skeletal muscle satellite cells real-time fluorescent quantitative PCR internal reference gene screening, the steps are as follows:

[0054] Cultivate wild-type, bovine skeletal muscle satellite cells transfected with interference MSTN and control, extract RNA and reverse transcribe;

[0055] Select several candidate internal reference genes, perform fluorescent quantitative P...

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Abstract

The invention relates to a method for interfering real-time fluorescent quantitative PCR reference gene screening of MSTN bovine skeletal muscle satellite cells. The method comprises the following steps of culturing wild type and transfecting interfering MSTN and contrast bovine skeletal muscle satellite cells, extracting RNA, and carrying out reverse transcription; selecting a plurality of candidate reference genes, carrying out fluorescent quantitative PCR reaction, and making a standard curve; judging the amplification efficiency of primers and the linear relationship between the template concentration and a Ct value; and analyzing the expression stability of the candidate reference genes, and taking the candidate reference gene with the optimal expression stability as the screened reference gene. According to the method, a proper reference gene is researched and screened out for interfering gene expression of the MSTN bovine skeletal muscle satellite cells, a powerful guarantee isprovided for accuracy of fluorescent quantitative PCR detection for interfering gene expression of the MSTN bovine skeletal muscle satellite cells, and reference can be provided for gene expression analysis of the bovine skeletal muscle satellite cells in different treatment and different periods in the future.

Description

technical field [0001] The invention belongs to the technical fields of biology, cell and tissue engineering, in particular to a method for interfering with MSTN bovine skeletal muscle satellite cells for real-time fluorescent quantitative PCR internal reference gene screening. Background technique [0002] The development and formation of muscle needs to go through a complex multi-level process, which is related to the action of many factors, especially the regulation of genes related to muscle development. It has been reported in the literature that genes affecting muscle development mainly play two regulatory roles, positive promotion and negative inhibition, and the mechanism of action is very complicated. The expression of genes that affect muscle development regulates the structure and function of cells, thereby further affecting the growth and development of muscles, and the degree of muscle development is the most intuitive measure of the profit of the breeding indus...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6888C12Q2600/124C12Q2600/158C12Q2600/166C12Q2561/113C12Q2563/107C12Q2545/114
Inventor 郭益文盛辉张林林胡德宝李新丁向彬郭宏
Owner TIANJIN AGRICULTURE COLLEGE
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