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Gene probe for identifying ingredients of sheep origin and goat origin

A technology of genetic probes and derived components, which is applied in the determination/testing of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., and can solve the problems of no probe design

Inactive Publication Date: 2013-11-27
新疆维吾尔自治区种羊与羊毛羊绒质量安全监督检验中心
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AI Technical Summary

Problems solved by technology

Subramanian et al. extracted the mitochondrial DNA in the fiber hair shaft, amplified the conserved region of mitochondrial DNA cytochrome b gene by PCR, and then selected the appropriate restriction site to analyze the single nucleotide polymorphism in the corresponding conserved region. The polymorphism of the fragment length after enzyme digestion can identify the source of fiber, so as to quickly and accurately distinguish cashmere and sheep wool. Although this method can be used to identify the origin of goat and sheep, it uses enzyme digestion and has no designed probe. Needle

Method used

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  • Gene probe for identifying ingredients of sheep origin and goat origin
  • Gene probe for identifying ingredients of sheep origin and goat origin
  • Gene probe for identifying ingredients of sheep origin and goat origin

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Embodiment

[0035] Applied to the detection of sheep wool mixed with cashmere:

[0036] 1) The invention is a pair of gene probes:

[0037]

[0038] The probe is located on the cytochrome b gene (Cytb), and the general primers used to amplify this fragment are:

[0039] 5'-GAAGCTTGATATGAAAAACCATCGTTG-3'; 5'-AAACTGCAGCCCCCTCAGAATGATATTTGTCCTCA-3'.

[0040] 2) Cut the plush sample into pieces, rinse twice with absolute ethanol, and extract DNA by phenol-chloroform method or kit method.

[0041] 3) PCR amplification: The primers refer to the primers for amplifying the mtDNA Cytb gene in Zhang Chuansheng's "Journal of Animal Husbandry and Veterinary Medicine" in 2005. The amplification range is located in the mtDNA genome 14143bp-14517bp, Cytb gene (-16bp-398bp), and the length of the amplified fragment is about 415bp .

[0042] The primer is: 5′-AAAAAGCTTCCATCCAACATCTCAGCATGATGAAA-3′;

[0043] 5'-AAACTGCAGCCCCTCAGAATGATATTTGTCCTCA-3'.

[0044] PCR reaction system: 10×PCR buffer 5μL, ...

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Abstract

The invention provides a gene probe for identifying ingredients of sheep origin and goat origin, which comprises a pair of designed gene probes, that is a sheep origin probe 5'-CTGTAACCCACATTTGCCGAGACG-3'bp24 and a goat origin probe 5'-TGATGAGATGTTTGATGGGG-3'bp20; the probes are positioned on a cytochrome gene b, that is Cytb; used primers are 5'-AAAAAGCTTCCATCCAACATCTCAGCATGATGAAA-3' and 5'-AAACTGCAGCCCCTCAGAATGATATTTGTCCTCA-3'; the amplified fragment is 530-550 bp. Through the adoption of a phenol- chloroform method or a kit method, the DNA of a to-be-tested sample is extracted, PCR amplification is implemented, a point system chip is designed, and hybridization, cleaning and the like are carried out.

Description

technical field [0001] The invention relates to the identification of goat and sheep-derived components, and is suitable for identifying sheep and goat-derived components by gene chip method, and is suitable for some tissues that are difficult to extract nuclear DNA or extract mitochondrial DNA (mtDNA), such as plush fibers; the probe is located in Universal primers amplify the range, which is convenient and fast to identify. Background technique [0002] In daily life, due to the advantages of goat and sheep products, there are price differences in meat, milk products, hides and plush fibers from goats or sheep. For example, the price of cashmere is much higher than that of sheep wool, so In order to reduce costs, some producers mix sheep wool or other fibers similar to cashmere into fabrics, so that counterfeit or reduced cashmere textiles appear on the market, which greatly damages the interests of consumers and damages sheep, goats, etc. Product quality, market circulat...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 郑文新高维明吕雪峰阿不力孜帕娜尔段新华陶卫东王乐杨会国宫平郑天健何茜邢巍婷张敏周卫东采复拉胡波隋恒风乌兰巍佩玲师帅左晓佳柴婷
Owner 新疆维吾尔自治区种羊与羊毛羊绒质量安全监督检验中心
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