A method for identifying sheep meat production traits and special primers

A technology for sheep and traits, applied in biochemical equipment and methods, microbe measurement/inspection, DNA/RNA fragments, etc., to achieve the effects of low cost, improved economic benefits, and simple operation

Active Publication Date: 2022-06-14
SHIHEZI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although our country is a big sheep-raising country in the world, our country is not a strong sheep-raising country. Therefore, the production capacity of mutton in our country needs to be improved

Method used

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  • A method for identifying sheep meat production traits and special primers
  • A method for identifying sheep meat production traits and special primers

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Determination of embodiment 1 SNP site

[0021] 1. Selection of experimental materials

[0022] Collect 40 muscle tissues of Hu sheep and 39 muscle tissues of Suffolk sheep (Xinao Animal Husbandry Co., Ltd., Manas County, Changji, Xinjiang), and extract genomic DNA respectively, and adjust the DNA concentration of each sample to 100ng / μL, a total of 40 DNA samples of Hu sheep and 39 DNA samples of Suffolk sheep were obtained, which were stored at -80°C for future use.

[0023] 2. Design and synthesis of primers

[0024] Since the mature sequence of the sheep miR-1 gene has not been published, the mature sequence of the bovine miR-1 gene (MIMAT0009214, ID: bta-miR-1) was retrieved from the miRBase database (http: / / www.mirbase.org / ) , and compared with the NCBI sheep genome database (http: / / www.ncbi.nlm.nih.gov), and intercepted the 200bp sequences on the left and right flanks, and designed a pair of primers (primer combination) to clone sheep miR-1 Precursor gene, the...

Embodiment example 2m

[0035] Implementation case 2 Establishment of asymmetric PCR-SSCP detection method for miR-1 precursor gene

[0036] 1. PCR amplification

[0037] The genomic DNA of the sheep to be tested obtained in step 1 was used as a template, and U and D were used as primers to carry out PCR amplification to obtain PCR amplification products. PCR amplification system: 1 μL of 100ng genomic DNA, 10 μL of 2×Taq PCR MasterMix, 0.5 μL of forward primer (concentration of 10 μmol / L), 0.5 μL of reverse primer (concentration of 10 μmol / L), add ddH2O to 20 μL. PCR amplification program: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 30 s; annealing at 60°C for 30 s; extension at 72°C for 50 s, a total of 30 cycles; final extension at 72°C for 10 min, detected by 1% agarose gel electrophoresis.

[0038] 2. Asymmetric PCR amplification conditions

[0039] The second PCR method is used, that is, the first PCR amplification is performed with primers of equal concentration to amplify t...

Embodiment example 3

[0045] Implementation Case 3 Correlation Analysis of Sheep miR-1 Precursor Gene g.53971777A>G Polymorphism and Meat Production-related Traits of Sheep

[0046] In order to determine whether the g.53971777A>G polymorphism site is related to the meat production of sheep, 40 Suffolk sheep and 39 Hu sheep were used as experimental materials, their genotypes were determined, and the relationship between the gene mutation site and the meat production Quantitative correlation analysis. Using SPSS13.0 software to calculate the genotype and allele frequency of the g.53971777A>G polymorphism site in the sheep miR-1 precursor gene in the population, and use the Chi-square test to analyze whether the genotype in each population is in the Hardy- Weinberg balance. The results are shown in Table 1.

[0047] The distribution and Hardy-Weinberg equilibrium of the gene frequency and genotype frequency of the SNP site of miR-1 gene in various sheep breeds

[0048]

[0049] Note: The number...

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Abstract

The invention discloses a method and application for assisting identification of sheep meat production-related traits. The invention provides a method for auxiliary identification of sheep meat-related traits. The specific steps are: using specific primers to perform PCR amplification on the genomic DNA of the sheep to be tested, and using DNA sequencing and asymmetric PCR-SSCP technology to detect DNA polymorphism And rapid genotyping, detect that the 209th nucleotide from the 5' end of the PCR amplification product is A or G, and determine the genotype of the sheep to be tested as GG or AA, and the SNP site is related to sheep meat production traits There is a certain correlation between them, and the meat production performance of GG genotype sheep is higher than that of AA genotype sheep. The invention can carry out early screening of sheep meat production, effectively increase the meat production of cultured sheep, reduce breeding costs, and improve the economic benefits of cultured sheep. The method of the invention is simple in operation, low in cost and high in accuracy, and has great significance for sheep breeding.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method and application for assisting in identifying traits related to sheep meat production. Background technique [0002] Sheep play an important role in both agricultural production and people's life, and are important livestock animals. Over the years, the number of sheep and goats in my country, the production of mutton, sheepskin, sheep casings and cashmere have ranked first in the world. At the same time, the price of domestic mutton has always been higher than that of pork, beef and chicken, and has been rising all the way. With the improvement of people's living standards, the types of meat on the table are also increasing, and people are no longer simply satisfied with pork and other meats with high fat content and high cholesterol content. The protein and essential amino acid content of sheep meat is higher than that of pork, chicken and beef, the fat conten...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156C12Q2600/124
Inventor 胡圣伟倪伟张翔宇李村院李晓悦
Owner SHIHEZI UNIVERSITY
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