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Simplified gene sequencing method of Peking duck based on gbs technology

A technology of gene sequencing and Peking duck, applied in the fields of molecular biology and high-throughput sequencing, can solve the problems of high price and low quality of reference genome, and achieve the effect of low price, low sequence redundancy and good specificity

Active Publication Date: 2019-04-16
CHINA AGRI UNIV
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AI Technical Summary

Problems solved by technology

However, the research on Peking duck has always been limited by the difficulty of genome information determination, because ducks lack a corresponding gene chip measurement platform, and the whole gene resequencing of Peking duck has the disadvantages of high price and low quality of reference genome

Method used

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  • Simplified gene sequencing method of Peking duck based on gbs technology
  • Simplified gene sequencing method of Peking duck based on gbs technology
  • Simplified gene sequencing method of Peking duck based on gbs technology

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Embodiment 1

[0027] Example 1 Peking Duck Simplified Gene Sequencing and Genotyping Method Based on GBS Technology

[0028] 1. GBS genotyping of Peking duck

[0029] 1. Select 50 6-week-old Peking ducks, collect 3ml of blood by tibial vein blood collection, and extract genomic DNA by traditional phenol imitation method, ddH 2 O was dissolved, and the genome integrity was confirmed by gel electrophoresis, and stored at -20°C for later use.

[0030] 2. Select an appropriate amount of genomic DNA that has passed the quality inspection, and use the Qiagen DNA purification kit to perform column purification to remove impurities such as protein, sugar, and salt, so as to prevent impurities from affecting the enzyme digestion efficiency, and use Qubit for accurate quantification of the purified product. To control the digestion time.

[0031] 3. Take 100ng of purified genomic DNA and fully digest it with MseI. Digestion system (50μL): 1μL restriction endonuclease MseI (100000U / mL), 100ng DNA, 5...

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Abstract

The invention provides a simplified Peking duck gene sequencing method based on a GBS technology. The method comprises the steps that Peking duck genome DNA is extracted, digestion is conducted on the genome DNA by means of restriction endonuclease MseI, digested fragments are recycled, specific joint sequences as shown in SEQ ID NO:1-12 are added, the digested fragments which have the size ranging from 550 bp to 580 bp and are provided with the joint sequences are selected for constructing a GBS library, quality control is conducted on the library, high-throughput sequencing is conducted, a sequencing result is compared to a duck reference genome, quality control is conducted on a comparison result, and Peking duck genome information is obtained finally. The method is easy to operate, accurate in result and suitable for actual breeding work of Peking ducks.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and high-throughput sequencing, in particular to a simplified gene sequencing method of Peking duck based on GBS technology. Background technique [0002] Peking duck is one of the agricultural animal varieties with great economic value in my country, and has a broad consumer market in the world, especially in Asia (Zhu F., Yuan J.M., Zhang Z.H., Hao J.P., Yang Y.Z., Hu S.Q., Yang F.X., Qu L.J. & Hou Z.C. (2015) De novo transcriptome assembly and identification of genes associated with feed conversion ratio and breast muscle yield in domestic ducks. Anim Genet 46, 636-45). At the same time, Peking duck is also a high-quality experimental material in the field of population genetics and biological evolution (Li H.F., Zhu W.Q., Song W.T., Shu J.T., Han W. & ChenK.W. (2010) Origin and genetic diversity of Chinese domestic ducks. MolPhylogenet Evol 57,634 -40; Hou Z.C., Yang F.X., Qu L.J., ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6869C12N15/11
CPCC12Q1/6869C12Q2525/191C12Q2521/301
Inventor 侯卓成朱峰崔倩倩
Owner CHINA AGRI UNIV
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