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Method for detecting single nucleotide polymorphism of sheep PCNP (PEST-Containing Nuclear Protein) gene by using PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism) and application of method

A single nucleotide polymorphism, sheep technology, applied in the field of molecular genetics, can solve problems such as lack of research, achieve the effect of improving selection efficiency, simple method, and speeding up breeding progress

Active Publication Date: 2017-05-31
甘肃润牧生物工程有限责任公司
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Problems solved by technology

Due to the lack of research on the genetic variation of the PCNP gene in Hu sheep and small-tailed Han sheep in China, the research on the function of the gene locus and the relationship between the genetic variation of the gene and economic traits, such as reproductive traits, have become blank

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  • Method for detecting single nucleotide polymorphism of sheep PCNP (PEST-Containing Nuclear Protein) gene by using PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism) and application of method
  • Method for detecting single nucleotide polymorphism of sheep PCNP (PEST-Containing Nuclear Protein) gene by using PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism) and application of method
  • Method for detecting single nucleotide polymorphism of sheep PCNP (PEST-Containing Nuclear Protein) gene by using PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism) and application of method

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Embodiment Construction

[0033] The present invention will be described in further detail below in conjunction with the accompanying drawings and embodiments, and the described embodiments are explanations of the present invention, rather than limitations.

[0034] The invention uses the conserved sequence of the PCNP gene to design primers to amplify the 686bp fragment of the second exon of the PCNP gene, and uses the genomes of Hu sheep and Small-tailed Han sheep as templates to perform PCR amplification. Nucleotide polymorphism: To conduct trait correlation analysis for the discovered single nucleotide polymorphism, and provide its detection method, making the nucleotide polymorphism of the PCNP gene a molecular genetic marker that can be quickly and conveniently detected, It provides a basis for accelerating the establishment of sheep populations such as Hu sheep and small-tailed Han sheep with high-quality economic traits.

[0035] a. Detection of PCNP gene polymorphism in sheep

[0036] 1. Coll...

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Abstract

The invention discloses a method for detecting single nucleotide polymorphism of a sheep PCNP (PEST-Containing Nuclear Protein) gene by using PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism) and application of the method. The method comprises the following steps: carrying out PCR amplification on a sheep PCNP gene segment by taking a sheep whole genome DNA to be measured as a template and taking a primer pair P as a primer; after a PCR amplification product is digested by a restriction enzyme SPhI, carrying out agarose gel electrophoresis on the digested amplified segment; authenticating the single nucleotide polymorphism of the 5019th site of the sheep PCNP gene according to an electrophoresis result. As a base mutation site is closely associated with the kidding number of sheep reproduction traits, the method is a method for detecting molecular genetic markers closely associated with the sheep reproduction traits at the DNA level, and can be used for auxiliary selection and molecular breeding of sheep and increasing the improved variety breeding speed of the sheep.

Description

technical field [0001] The invention belongs to the field of molecular genetics, relates to using the single nucleotide polymorphism (SNP) of sheep functional gene as a molecular genetic marker, in particular to the single nucleotide polymorphism of sheep PCNP gene and a detection method thereof. Background technique [0002] In sheep breeding, it is expected that through the selection of DNA markers that are closely related to reproductive traits and closely linked to quantitative traits, the purpose of early selection and improvement of the accuracy of breeding values ​​can be achieved, so as to obtain greater heredity in livestock and poultry breeding. progress. [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 tradition...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/683C12Q2531/113C12Q2521/301C12Q2565/125
Inventor 乐祥鹏曹学涛孙武李发弟王维民李万宏
Owner 甘肃润牧生物工程有限责任公司
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