A method for screening high-quality black black cattle breeds and the primers and kits used

Abstract

The invention discloses a method for screening high-quality bulls of Blackett black cattle. The method comprises the steps that a gene sequence of the second exon of a PID1 gene of the Blackett black cattle is subjected to PCR amplification, and the Blackett black cattle with the base at the 285th position of the second exon of the PID1 gene being G are selected as the high-quality bulls. The invention further discloses a primer and a kit demanded by screening. The method for screening the high-quality bulls by detecting PID1 gene base mutation is raised for the first time, and a new idea is provided for cultivating high-quality Blackett beef cattle breeds. A PCR-RFLP method is used preferentially to detect base mutation and genotype of the Blackett black cattle, the high-quality bulls can be screened out through the number of electrophoresis fragments, a sequencing technology for SNP screening and the PCR-RFLP technology are combined, and accordingly defects of instability of SSCP and limitations of ordinary RFLP are overcome; the method is simple and quick, cost is low, precision is high, detection results are visual, and popularization is facilitated.

Description

Technical field [0001] The invention relates to a method for screening high-quality breeds of Black Cattle cattle, a primer used for amplifying the second exon of PID1 gene, and a kit used for screening high-quality breeds, belonging to the technical field of animal molecular genetics. Background technique [0002] With the enrichment of food culture and the continuous improvement of people's living standards, more and more people are aware of the unique nutritional value of beef. Beef contains essential amino acids needed by the human body, has the characteristics of high protein, low cholesterol, and rich nutrition. With the increase of consumer groups, the proportion of beef in meat consumption has been rising year by year, and beef prices have continued to rise. While the total consumption is increasing, people have higher requirements for the quality of beef. "Snow beef" and "high-end beef" have gradually become fashionable consumption, and consumers' demand for high-end be...

AI Technical Summary

Problems solved by technology

[0004] At present, there are few reports on methods for screening or cultivating high-quality Black Cattle breeds. At present, SSCP and common RFLP techniques are commonly used for single nucleotide polymorphisms (Single nucleotide polymorphisms, SNPs) detection and population Genetic polymorphism analysis, these methods are unstable, some mutations do not cause DNA bending, and molecular conformation does not change. Under normal conditions, these mutations may not be detected. Therefore, these techniques alone cannot prove that there is no mutation at all. In actual application, a considerable proportion of false negative results may be encountered; meanwhile, the sensitivity of the current inspection technique is not high, which is affected by various factors such as electrophoresis temperature, ionic strength (ie, the concentration of the electrophoresis buffer), gel concentration, glycerol concentration and interaction. The influence of the concentration of the coupling agent methylenebisacrylamide, etc., leads to poor repeatability; in addition, its operation process is complicated, which greatly limits its application in production

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