Method for detecting molecular marks of butter-fat content of Chinese Holstein by utilizing LDR technology

Abstract

The invention discloses a method for detecting molecular marks of the butter-fat content of Chinese Holstein by utilizing the ligase detection reaction (LDR) technology, which is characterized by comprising the following steps of: (1) performing amplification on a fragment which is used for an LDR reaction and contains A8398G loci, namely by taking deoxyribonucleic acid (DNA) of an A8398G genome of the Chinese Holstein as a template, performing the amplification by using a primer under the condition of a polymerase chain reaction (PCR) in the presence of thermus aquaticus (Taq) DNA polymerase, buffering environment, Mg<2+> and dNTPs and using the product of the amplification for the LDR reaction; (2) performing the LDR reaction, namely designing three probes, and performing the LDR reaction; (3) analyzing, namely detecting an LDR reaction product by using an ABI PRISM-377 DNA sequencing instrument, and performing correlation analysis according to the length of the LDR product and eachgenotype. In the method for detecting the molecular marks of the butter-fat content of the Chinese Holstein by utilizing the LDR technology, on the basis of basic group mispairing, the DNA of the specific fragments is identified by using the specific probes, so the method has the high accuracy; and compared with other single nucleotide polymorphism (SNP) detection technologies, the LDR detection technology has the advantages of high accuracy and generality, large flux, simplicity of operation, low cost and the like.

Description

technical field [0001] The invention relates to a method for detecting molecular markers of milk fat content of Chinese Hesiyuan cattle by using LDR technology, which belongs to the field of molecular genetics. technical background [0002] Milk production traits are economically important traits in the dairy industry. The traditional view of quantitative genetics holds that the milk production traits of dairy cows (including milk yield, milk fat, protein content, milk fat percentage, milk protein percentage, etc.) are quantitative traits, determined by micro-effect polygenes, and affected by environmental factors. Since the effect of a single minor gene cannot be directly observed and measured, statistical genetics can only be used to describe the genetic nature of quantitative traits. This research method has played a huge role in the genetic improvement of animals, but it is impossible to completely eliminate environmental factors, and there are certain deviations. [0...

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Problems solved by technology

[0005] At present, the methods for detecting DNA mutation sites generally use restriction fragment length polymorphism (Restriction Fragment Length Polymorphism, RFLP) technology and single-strand conformational polymorphism (Single-Strand Con-formational Polymorphism, SSCP) technology, these two methods rely on the naked eye. Observation of the results, sometimes there will be errors in judgment, which will affect the reliability of the test results

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