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Screening method of reference genes for fluorescent quantitative detection of caruncle mucosa of pregnant cows and application of screening method

A technology of fluorescent quantitative detection and internal reference genes, which is applied in the determination/testing of microorganisms, biochemical equipment and methods, etc., and can solve the problems of unconfirmed expression stability

Pending Publication Date: 2019-06-21
WUHAN ACADEMY OF AGRI SCI
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Problems solved by technology

However, the expression stability of commonly used internal reference genes in the fluorescence quantitative detection of pregnant bovine caruncle mucosa tissue has not yet been confirmed

Method used

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  • Screening method of reference genes for fluorescent quantitative detection of caruncle mucosa of pregnant cows and application of screening method
  • Screening method of reference genes for fluorescent quantitative detection of caruncle mucosa of pregnant cows and application of screening method
  • Screening method of reference genes for fluorescent quantitative detection of caruncle mucosa of pregnant cows and application of screening method

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Embodiment Construction

[0037] The present invention will be further described in detail below in conjunction with the accompanying drawings and specific embodiments.

[0038] The embodiment of the present invention provides a method for screening internal reference genes of pregnant bovine caruncle mucous membrane fluorescent quantitative detection, comprising the following steps:

[0039] S1, extracting the RNA of the caruncle mucosa tissue, and performing reverse transcription;

[0040] (1) Pretreatment of experimental animals:

[0041] The 3 test cows came from a standardized dairy cow demonstration farm in Wuhan, with normal reproductive system and fertility, and they were all multi-parity cows. Estrus synchronous drugs follicle-stimulating hormone (FSH), loprostinol (PG) and injection of gonadorelin (GnRH) were purchased from Ningbo Sansheng Pharmaceutical Co., Ltd. On the 9th day of the estrous cycle of the test cows, superovulation treatment was carried out, and follicle-stimulating hormone...

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Abstract

The invention discloses screening method of reference genes for fluorescent quantitative detection of caruncle mucosa of pregnant cows and application of the screening method, and relates to the technical field of animal molecular biology. Suitable reference genes are screened out for the study on gene expression of caruncle mucosa of pregnant cows; the expression of genes playing a key role uponestablishing of a bond between the maternal cow and fetus can be truly reflected; key candidate genes of maternal-fetal interaction can be further applied to assist in the important reproductive mechanisms, such as cow pregnancy establishment, pregnancy recognition, and pregnancy maintenance.

Description

technical field [0001] The invention relates to the technical field of animal molecular biology, in particular to an internal reference gene screening method for quantitative detection of pregnant bovine caruncle mucous membrane fluorescence and its application. Background technique [0002] Fluorescent quantitative PCR is a method for quantifying mRNA expression at the transcription level. Compared with conventional PCR, it has the advantages of high sensitivity, good specificity, high accuracy, fast speed and high throughput. Common means of expression and molecular function. When using fluorescent quantitative PCR technology to detect the expression level of the target gene, in order to reduce the possible errors caused by the RNA yield, quality and reverse transcription efficiency of the test sample, stable internal reference genes are usually used for correction and standardization. [0003] A large number of studies have confirmed that the expression of the same inter...

Claims

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Application Information

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IPC IPC(8): C12Q1/6811C12Q1/6851
Inventor 程蕾向敏夏瑜陶弼菲余婕胡修忠王定发周源
Owner WUHAN ACADEMY OF AGRI SCI
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