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Method for detecting egg fishy odour mutation allele and kit thereof

A technology of alleles and fishy smell, applied in the field of epitopes, can solve the problems of difficult breeding, many detection steps, and easy occurrence of miscellaneous bands, etc., and achieve the effects of saving feed costs, simple operation, and low cost

Active Publication Date: 2008-07-09
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The PCR-SSCP method is the most commonly used method, but the existing problems are: more detection steps; longer time-consuming (12-20 hours); easy to appear miscellaneous bands, increase the probability of misjudgment, and the reliability of the results is low
Although DNA sequencing, allele-specific oligonucleotide hybridization, DNA chips and TaqMan methods are relatively high in accuracy, they are expensive to detect and difficult to apply to actual breeding

Method used

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  • Method for detecting egg fishy odour mutation allele and kit thereof
  • Method for detecting egg fishy odour mutation allele and kit thereof

Examples

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Comparison scheme
Effect test

Embodiment 1

[0026] Example 1. Establishment of a method for detecting fishy smell mutant alleles in eggs

[0027] 1. Determination of the A985T polymorphic site

[0028] 1. PCR amplification

[0029] The pure line of dwarf brown-shell hens was selected as the experimental material. Primers were designed according to GenBank Accession Number AJ431390 so that the amplified target fragment contained the 1034th deoxyribonucleotide from the 5' end of GenBank Accession Number AJ431390. The primer sequences were as follows:

[0030] U (upstream primer): 5'-CAGGGTGGTATCAAGCCTGT-3' (sequence 1 in the sequence listing)

[0031] D (downstream primer): 5'-GATCCAAAGGACTGGACCAA-3' (sequence 2 in the sequence listing)

[0032] Using the pure line genomic DNA of dwarf brown-shell hens as a template, PCR amplification was carried out using primers U and D.

[0033] The amplification system is as follows:

[0034] Chicken Genomic DNA 50ng

[0035] 10×PCR Amplification Buffer 1.5μL

[0036] dNTPs (4mmo...

Embodiment 2

[0070] Example 2. Detection kits for detection of fishy smell mutant alleles in eggs of different breeds of chickens carrying fishy smell mutant alleles in eggs and genotype-correlation verification analysis of fishy smell egg traits produced by chickens

[0071] Feeds containing rapeseed (containing trimethylamine) were used to feed white legion hens, H brown hens, and Dongxiang green-shell hens.

[0072] The A985T polymorphism of the above-mentioned different breeds of chickens was detected by using the egg fishy smell mutant allele detection kit. The egg fishy smell mutant allele detection kit includes: a pair of primers, wherein the nucleotide sequence of one primer is sequence 1 in the sequence listing, and the nucleotide sequence of the other primer is sequence 2 in the sequence listing; restriction nucleic acid Endonuclease BsrI; the kit also includes routine reagents required for PCR-RFLP. Using the genomic DNA of the chicken to be tested as a template, perform PCR am...

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Abstract

The invention discloses a process of detecting mutation allelomorph of egg fishy odor and an agent case, which can be applied in screening the property of a chicken which lays egg with fishy odor. Chickens whether carry mutation allelomorph of egg fishy odor can be accurately and high effectively differentiated through using the process or the agent case of the invention, thereby solving the problem of egg fishy odor in practical production, greatly reducing the cost of molecule breeding, being beneficial for widespread choosing feed raw material, reducing feed costs and increasing economic benefits. The detecting process or the agent case which are provided by the invention has simple operation, low cost, short detecting time and high accuracy, which can realize automatic detection, and will plays great role in chicken breeding.

Description

technical field [0001] The invention relates to a method and a kit for detecting mutant alleles of fishy smell in eggs. Background technique [0002] With the development of the economy and the improvement of material living standards, people have higher and higher requirements for the quality of livestock products. The demand for eggs is no longer limited to quantity, but also has higher requirements for quality and flavor. Usually fishmeal in feed will cause chickens to produce fishy-smelling eggs. However, some chickens still produce fishy-smelling eggs when fed with feed that does not contain fishmeal. Because the egg must be opened to determine whether it has a fishy smell, it has caused great economic losses to poultry producers and processors. Therefore, it is of great economic and social significance to study the mechanism of the fishy smell of eggs and eliminate it through breeding. [0003] According to the research of Bolton et al. (1976), it was found that the ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 杨宁张龙超郑江霞
Owner CHINA AGRI UNIV
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