Method for preparing single embryo chromosome of flounder fish

A chromosome and embryo technology, applied in the field of marine animal cytogenetics, can solve the problems of impact analysis, less fission phase, material waste, etc., and achieve good results without wasting materials

CN103267665AInactive Publication Date: 2013-08-28OCEAN UNIV OF CHINA
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Publication Date
2013-08-28
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention relates to a method for preparing a single embryo chromosome of flounder fish and belongs to the field of marine animal cytogenetics in biotechnology. The embryo of the flounder fish is used as a material, an injector is used as a tool, the egg membrane of the embryo material is broken by means of physical pressurization, the embryo between tail germ stage and hatching stage is placed in a syringe together with seawater, the piston of the injector is pushed so that the embryo is extruded out of the needle part, and then the membrane of the embryo can be broken and the tissue can be preserved relatively completely, and the chromosome preparation of the membrane-broken embryo is carried out according to a conventional chromosome preparation method; and the chromosome preparation specifically comprises the following steps of treating the membrane-broken embryo for two hours by using colchicine, then treating in a hypotonic solution for 30 minutes, and at last fixing the embryo by using a Carnot fixing solution, wherein each Carnot fixing takes 20 minutes. A lot of chromosome dividing phases of the flounder fish with very good effect are obtained by using the method provided by the invention, especially the chromosome of a single embryo can also be prepared without wasting material, and subsequent genetics analysis can be carried out on a single individual.
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Description

technical field

[0001] The invention belongs to the field of marine animal cytogenetics in biotechnology, and in particular relates to a chromosome preparation method suitable for a single embryo of flounder and flounder. Background technique

[0002] Flounder and plaice belong to the class Teleost ( Osteichthyes ), Flatfish ( Pleuronectiformes ), commonly known as flatfish ( Flatfishes ), can be divided into Flounderidae ( Bothidae ), Flounder ( Pleuronectidae ) and Soleidae ( Soleoidae ). Flounder is an important part of marine fish cultured in China. Its meat is delicious and tender, rich in nutrition, and has extremely high nutritional and economic value.

[0003] With the continuous development of marine vertebrate cytogenetics, the chromosome preparation technology of flounder and flounder has been relatively mature, and it is relatively convenient to prepare chromosomes of flounder and flounder by means of gill filaments, kidney or cell culture and carry ou...

Examples

Embodiment 1

[0013] Embodiment 1. Chromosome preparation of flounder embryos using the method of the present invention

[0014] Select the flounder that enters the filming stage ( Paralichthys olivaceus, Belonging to Plaiceiformes, Flounder family, Flounder genus) embryos are used as materials, which are placed in a syringe together with the seawater for incubation, and then the piston of the syringe is pushed to squeeze the embryos out from the needle. Embryos after membrane rupture were treated in 0.005% (mass volume ratio) colchicine for 2 hours, then placed in 0.075mol / L KCl hypotonic solution for 30 minutes, and finally Carnot's fixative (ethanol: glacial acetic acid = 3:1, volume ratio) fixed 3 times, 20min each time.