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Method for detecting single nucleotide polymorphism of sheep DAQL gene by using PCR-RFLP and application of method

A single nucleotide polymorphism and detection method technology, which is applied in the field of detection of single nucleotide polymorphism of sheep DAZL gene, can solve the problems of cumbersome operation and high cost, and achieve the purpose of solving false positives, improving accuracy and saving genes The effect of the typing step

Active Publication Date: 2019-12-20
甘肃润牧生物工程有限责任公司
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AI Technical Summary

Problems solved by technology

The RFLP-PCR method not only has the accuracy of DNA sequencing, but also overcomes the shortcomings of expensive, cumbersome operations, and false positives, and there are no special requirements for the sequence sites detected

Method used

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  • Method for detecting single nucleotide polymorphism of sheep DAQL gene by using PCR-RFLP and application of method
  • Method for detecting single nucleotide polymorphism of sheep DAQL gene by using PCR-RFLP and application of method
  • Method for detecting single nucleotide polymorphism of sheep DAQL gene by using PCR-RFLP and application of method

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Embodiment 1

[0053] The method for detecting the single nucleotide polymorphism of sheep DAZL gene by PCR-RFLP includes the detection method of detecting the single nucleotide polymorphism of sheep DAZL gene whose 26750th position is G or A by PCR-RFLP, said detection method Include the following steps:

[0054] S1 Collect the testis tissue from the slaughtered Hu sheep testis, store it in a liquid nitrogen environment, and then grind it into powder. Take 0.1g of the obtained powder and place it in 1000μL DNA extraction buffer, and treat it with proteinase K at 55°C. After being treated with proteinase K at 55°C, the solution after the reaction was treated with Tris-saturated phenol to obtain a DNA extract. The specific steps are:

[0055] a. Add an appropriate amount of liquid nitrogen to the mortar of the testicular tissue frozen in liquid nitrogen environment and undergo DEPC enzyme inactivation and high temperature sterilization, grind it into powder, take 0.1g and add it to 1000 μL DN...

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Abstract

The invention discloses a method for detecting a single nucleotide polymorphism of sheep DAQL gene by using PCR-RFLP, and particularly relates to a method for detecting a single nucleotide polymorphism of sheep DAQL gene with amino acid at 26750th position being G or A by using PCR-RFLP. The method comprises the following steps: S1, collecting a sample and performing early-stage treatment on the sample; S2, constructing a variety DNA pool; S3, treating the sample by a PCR-RFLP method; and S4, gel imaging to determine genotype; and two-way sequencing to determine the single nucleotide polymorphism. The invention also provides an application of the method for detecting single nucleotide polymorphism of sheep DAQL gene by using PCR-RFLP. The application comprises the following steps: I analysis of phenotypic data and statistical analysis of single nucleotide loci; II establishing a mathematical model through association analysis; and III constructing a model to derive the single nucleotide polymorphism of the DAQL gene in adult sheep. The method and the application have beneficial effects that: the variety DNA pool is treated by PCR-RFLP to achieve a purpose of accurate labeling; digestion with AluI enzyme is used to solve a problem of false positives; and the mathematical model is used for correlation analysis to improve accuracy of molecular markers.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to a method for detecting the single nucleotide polymorphism of sheep DAZL gene by PCR-RFLP and its application. Background technique [0002] In animal breeding, people expect to achieve early selection and improve the accuracy of breeding values ​​by selecting DNA markers that are closely related to growth, reproduction and other traits and are closely linked to quantitative traits, so as to obtain better results in livestock and poultry breeding. Great genetic advances. [0003] Molecular breeding, that is, Molecular Mark-Assist Selection (MAS), is a technology that uses DNA molecular markers to select genetic resources or breeding materials, and to improve the comprehensive traits of livestock and poultry. It uses modern molecular The method of combining biology and traditional genetic breeding is used to select and breed new varieties. [0004] Gene polymorphism refers to the di...

Claims

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Application Information

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IPC IPC(8): C12Q1/683G16B5/00
CPCC12Q1/683G16B5/00C12Q2531/113C12Q2521/301Y02P60/87
Inventor 乐祥鹏罗静秦芳李发弟李万宏
Owner 甘肃润牧生物工程有限责任公司
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