Application and detection method of single-stranded nucleotide sequence of inha gene as animal superovulation molecular marker

A single-stranded nucleotide and superovulation technology, which is applied to the application and detection field of animal superovulation molecular markers, can solve the problems of large sample consumption, large workload and high cost, and achieves convenient implementation, improved efficiency, and improved methods. easy effect

Active Publication Date: 2017-01-04
JILIN UNIV
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Problems solved by technology

However, RFLP analysis requires high sample purity, a large amount of samples, and low polymorphic information content of RFLP. The level of polymorphism depends too much on the type and quantity of restriction endonucleases. In addition, RFLP analysis technology steps are cumbersome and the workload is large. , high cost, so its application is limited

Method used

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  • Application and detection method of single-stranded nucleotide sequence of inha gene as animal superovulation molecular marker
  • Application and detection method of single-stranded nucleotide sequence of inha gene as animal superovulation molecular marker
  • Application and detection method of single-stranded nucleotide sequence of inha gene as animal superovulation molecular marker

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Experimental program
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Embodiment 1

[0050] 1. Genomic DNA was extracted from the blood of Xiangxi yellow cattle with the Genomic DNA Extraction Kit of Tiangen Biochemical Technology Co., Ltd.

[0051] 2. Partial fragment amplification of INHA gene of Xiangxi yellow cattle:

[0052] Primers:

[0053]

[0054] PCR process:

[0055]

[0056]

[0057] For PCR results, see figure 1 .

[0058] 3. SSCP typing of PCR amplification products of different individuals:

[0059] Mix 3 μl of PCR product with 7 μl of denaturing buffer (98% deionized formamide, 10 mmol / L EDTA (pH8.0), 0.025% xylene cyanol FF, 0.025% bromophenol blue), and use a PCR instrument 95 Denature at ℃ for 10 minutes, and then stand on ice for 5 minutes to avoid renaturation of the denatured product, thereby obtaining denatured single-stranded DNA. The denatured DNA was electrophoresed in a 12% non-denaturing polyacrylamide gel at 140V in a refrigerator at 4°C for 14h. The gel was silver-stained with 0.4% silver nitrate to develop the colo...

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Abstract

The invention relates to the field of biotechnology, and particularly relates to applications of a single-stranded nucleotide sequence of an INHA gene as animal superovulation molecule marker, and a detection method. The INHA gene of the western Hunan yellow cattle has Reference SNP Cluster Report:rs41257116 locus, and when being hybridized into A / G, the locus expresses the superovulation advantage. The partial segment at the Reference SNP Cluster Report:rs41257116 locus of the INHA gene of the to-be-detected western Hunan yellow cattle is detected by adopting a PCR-SSCP technology to determine the INHA gene of the western Hunan yellow cattle as AA or AG, the western Hunan yellow cattle with the INHA gene being AG type is selected as western Hunan yellow cattle capable of obtaining better superovulation character. According to the method, the superovulation efficiency of the western Hunan yellow cattle can be increased. The method is simple and convenient, fast and sensitive, a special instrument is not required, and the implementation is convenient.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application and detection method of the single-stranded nucleotide sequence of the INHA gene as an animal superovulation molecular marker. Background technique [0002] Since Smith and Engle first discovered the phenomenon of superovulation in the 1920s, superovulation technology has been applied to different mammals and played an important role. The use of superovulation technology can give full play to the reproductive potential of excellent female animals, provide more embryos for embryo transfer technology, and speed up breeding. In addition, superovulation technology is also used in cloning, embryo freezing, and production of transgenic animals. Xiangxi Yellow Cattle has the advantages of tolerance to rough feeding, strong disease resistance, good reproductive performance, and stable genetics. It is an excellent meat-serving local yellow cattle breed in Hunan Province. But...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q1/6888C12Q2600/156C12Q2531/113C12Q2565/133C12Q2565/125
Inventor 陈树雄李纯锦陈璐李万宏刘卓孙丽娜赵云李洪蛟于佳鑫周虚巴桑旺堆朱彦宾
Owner JILIN UNIV
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