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Method for identifying Chinese prawn population by using AFLP (Amplified Fragment Length Polymorphism) fingerprint technique

A technology of Chinese prawns and populations, which is applied in the fields of recombinant DNA technology, biochemical equipment and methods, and the determination/inspection of microorganisms to achieve the effect of safeguarding marine rights and interests

Inactive Publication Date: 2012-09-19
OCEAN UNIV OF CHINA
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Problems solved by technology

However, there is no effective molecular identification method to identify the affiliation of the Chinese shrimp population

Method used

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  • Method for identifying Chinese prawn population by using AFLP (Amplified Fragment Length Polymorphism) fingerprint technique
  • Method for identifying Chinese prawn population by using AFLP (Amplified Fragment Length Polymorphism) fingerprint technique
  • Method for identifying Chinese prawn population by using AFLP (Amplified Fragment Length Polymorphism) fingerprint technique

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Embodiment Construction

[0015] The experimental methods in the examples of the present invention that do not specify specific conditions can generally be operated under conventional conditions, such as the conditions described in the "Molecular Cloning Experiment Guide" written by J. Sambrook et al.

[0016] In the long-term research, the applicant has screened the combination of primers that can effectively identify the Chinese prawn population through the AFLP analysis of different populations, and uses the primer set of the present invention to identify the Chinese prawn population. The steps are as follows:

[0017] 1. Genomic DNA extraction

[0018] The whole genome DNA of Penaeus chinensis was extracted and stored at -20°C for future use.

[0019] 2. Enzyme digestion reaction

[0020] A total of 20 μL enzyme digestion reaction system: about 100 ng of genomic DNA, 0.1 μL Trul Ι (10u / μl), 0.1 μL CoR I (10u / μl) and 4.0 μL 10×Y+ / Tango buffer, add ultrapure water to 20 μL, and place in a constant t...

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Abstract

The invention relates to a primer set for identifying a Chinese prawn population. The primer set comprises four pairs of primers, wherein sequences of a forward primer and a reverse primer of each pair of primers are SEQ ID NO: 1 and SEQ ID NO: 2, SEQ ID NO: 1 and SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and SEQ ID NO: 6 and SEQ ID NO: 7 respectively. According to a primer combination screened by the invention, the source and the place of the Chinese prawn population at a natural sea area can be determined, and the contribution of different spawning sites to the Chinese prawn population is favorably defined. Chinese prawns are widely distributed in the sea area shared by China, Japan and Korea; according to the primer combination screened by the invention, the contribution of different spawning water areas to the condominium water area resource can be determined, and resource place proportion and deserved fishing quota can be defined; and the method has great significance to safeguard of Chinese maritime right and protection of Chinese fishery resources.

Description

technical field [0001] The invention belongs to the technical field of molecular detection of species group identification in aquatic products, and in particular relates to a method for identifying Chinese prawn populations by using AFLP fingerprint technology. Background technique: [0002] AFLP (Amplified Fragment Length Polymorphism, Amplified Length Polymorphism) is a new DNA molecular marker technology invented on the basis of RFLP and RAPD technology. The basic principle is to digest genomic DNA with restriction endonucleases to generate DNA fragments of different sizes, then connect them with double-stranded artificial adapters, and then use the complementary strands of the artificial adapters as primers for pre-amplification. The primers used in the selective amplification are to add 1-3 selective nucleotides on the basis of the complementary chain of the adapter. After the selective amplification of the pre-amplified template DNA, the selective amplification is dete...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 高天翔张辉张朝晖李鹏飞
Owner OCEAN UNIV OF CHINA
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