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Screening of reference genes for quantitative PCR (polymerase chain reaction) detection of bovine reproductive system and embryo tissues and application of screening

A technology of internal reference genes and reproductive systems, 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 real-time quantitative PCR detection of pregnant dairy cow reproductive system and embryonic tissue has not yet been confirmed.

Method used

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  • Screening of reference genes for quantitative PCR (polymerase chain reaction) detection of bovine reproductive system and embryo tissues and application of screening
  • Screening of reference genes for quantitative PCR (polymerase chain reaction) detection of bovine reproductive system and embryo tissues and application of screening
  • Screening of reference genes for quantitative PCR (polymerase chain reaction) detection of bovine reproductive system and embryo tissues and application of screening

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Embodiment Construction

[0043] The present invention will be further described in detail below in conjunction with the accompanying drawings and specific embodiments.

[0044] The embodiment of the present invention provides a method for screening internal reference genes by quantitative PCR detection of bovine reproductive system and embryonic tissue, comprising the following steps:

[0045] S1, extracting the RNA of the reproductive system and embryonic tissue of the pregnant dairy cow, and performing reverse transcription;

[0046] (1) Pretreatment of experimental animals:

[0047] 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...

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Abstract

The invention discloses screening of reference genes for quantitative PCR (polymerase chain reaction) detection of bovine reproductive system and embryo tissues and application of the screening. Suitable reference genes are screened out for the study on genetic expression of reproductive system and embryo tissues of pregnant cows; the accuracy of fluorescent quantitative PCR detection on the genetic expression of reproductive system and embryo tissues of pregnant cows is forcibly guaranteed; further study on the regulation mechanism of cow embryo development and the maternal-fetal interactionbasic law is facilitated; important theoretical support is provided for the deep understanding of related molecular mechanisms of pregnancy recognition and pregnancy establishment; theoretic basis isprovided for reducing early pregnancy failure by manual intervention means.

Description

technical field [0001] The invention relates to the technical field of animal molecular biology, in particular to screening and application of an internal reference gene for quantitative PCR detection of bovine reproductive system and embryo tissue. Background technique [0002] Fluorescent quantitative PCR technology is a common method for studying mRNA expression levels, and has the advantages of accurate quantification, high sensitivity, good repeatability, strong specificity, and batch operation. However, the accuracy and reproducibility of real-time PCR data analysis depends on the normalization method used. In gene expression analysis, an appropriate internal reference gene is usually selected to correct and standardize the target gene to improve the sensitivity and repeatability of the method. An ideal internal reference gene is one that can be stably expressed in various types of tissues or cells under various experimental conditions and influencing factors. Theref...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6809C12Q1/6851
Inventor 程蕾余婕王定发胡修忠向敏周源夏瑜陶弼菲
Owner WUHAN ACADEMY OF AGRI SCI
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