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SNP molecular markers related to chicken polydactyly and its application

A technology of molecular markers and traits, applied in the direction of recombinant DNA technology, DNA / RNA fragments, etc., can solve the problems of increasing the probability of misjudgment, high detection costs, and many detection steps, achieving considerable economic benefits, high accuracy, and simple operation Effect

Active Publication Date: 2018-02-27
CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The PCR-SSCP method is the most commonly used method, but there are problems: more detection steps; longer time-consuming (12-20 hours); miscellaneous bands are prone to appear, increasing 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 all relatively high in accuracy, the detection costs are high and it is difficult to apply them to actual breeding

Method used

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  • SNP molecular markers related to chicken polydactyly and its application
  • SNP molecular markers related to chicken polydactyly and its application
  • SNP molecular markers related to chicken polydactyly and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1. Establishment of a method for detecting polydactyly mutant alleles of Beijing oil chicken and silky chicken

[0029] 1. Determination of the C4645A polymorphic site

[0030] 1. PCR amplification

[0031] The pure line of Beijing Youji was chosen as the experimental material. Primers were designed according to GenBank AccessionNumber NC_006089, so that the amplified target fragment contained the 4645th deoxynucleotide of the 5th intron of GenBank AccessionNumber NC_006089, and the primer sequences were as follows:

[0032] U (upstream primer): 5'-GCGATTTCCTCTCACCCCACA-3' (SEQ ID No.2)

[0033] D (downstream primer): 5'-AGCTGAGCAACATGACAGCA-3' (SEQ ID No.3)

[0034] Using the genomic DNA of Beijing Youji pure line as a template, PCR amplification was carried out using primers U and D.

[0035] The amplification system is as follows:

[0036] Chicken Genomic DNA 50ng

[0037] 10X PCR Amplification Buffer 1.5 μm

[0038] dNTP (4mmol / L) mixed solution 0.8 μ...

Embodiment 2

[0070] Example 2 Detecting polydactyly mutant alleles carried by different breeds of chickens and genotype-chicken polydactyly trait correlation verification analysis by Beijing oil chicken and silky chicken polydactyly mutant allele detection kit

[0071] Beijing oil chicken, silky silky chicken and white Leghorn laying hens were selected as experimental materials.

[0072] The C4645A polymorphism of the above-mentioned different breeds of chickens was detected by using the Beijing oil chicken and silky chicken polydactyly mutant allele detection kit. The polydactyl mutation allele detection kit for Beijing oil chicken and silky chicken includes: a pair of primers, the nucleotide sequence of one primer is sequence 2 in the sequence list, and the nucleotide sequence of the other primer is sequence 2. Sequence 3 in the list; restriction endonuclease BsrDI; the kit also includes conventional reagents required by PCR-RFLP. Using the genomic DNA of the chicken to be tested as a t...

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Abstract

The invention provides a SNP molecular marker related to chicken polydactyly and primers for amplifying it. The molecular marker is from the LMBR1 gene, the nucleotide sequence of which is shown in SEQ ID No.1, and the 246th bp is a mutation site, the base is A or C. The invention also provides a method and a kit for detecting the polydactyly genotype of chickens. The detection method or kit provided by the invention is used to detect whether the Beijing oily chicken and the silky chicken carry the polydactyly mutant allele, and the polydactyly character of the Beijing oily chicken and the silky chicken can be determined. In actual production and breeding, polydactyly can be selected to be retained or eliminated according to needs, and the elimination effect is complete and accurate.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a SNP molecular marker related to chicken polydactyly and its application. Background technique [0002] Beijing oil chicken and silky silky chicken are famous for their unique appearance and delicious meat and eggs. They are precious and fine local breeds in my country. In natural populations, some individuals of Beijing oil chicken and silky chicken had polydactyly, but the polydactyly was not fixed in the population. In recent years, due to the complex and changeable natural environment, the frequency of outbreaks of various zoonotic diseases has become higher and higher, which not only caused human casualties, but also severely damaged the poultry breeding industry. Under this general environment, local governments Legislation has been enacted to abolish the trade in live poultry. However, the slaughtered Beijing oily chicken and silky silky chicken are difficult to dist...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 曲鲁江张泽宾陈余贾亚雄吕学泽陈继兰杨宁
Owner CHINA AGRI UNIV
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