Method for preparing chromosomes through large-scale barbel fish nephrocyte in-vivo culture

A large-scale barb carp, chromosome technology, applied in artificial cell constructs, animal cells, vertebrate cells, etc., to achieve the effect of clear shape, strong dyeability and good dispersion

Inactive Publication Date: 2013-05-08
HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no report on the in vivo culture

Method used

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  • Method for preparing chromosomes through large-scale barbel fish nephrocyte in-vivo culture

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specific Embodiment approach 1

[0020] Specific embodiment one: the method for preparing chromosomes in vivo of a kind of giant mackerel kidney cell of the present embodiment is carried out according to the following steps:

[0021] 1. In vivo injection: anesthetize the giant catfish with 5-10 ppm phenoxyethanol and weigh it, and inject phytohemagglutinin (PHA) into the intraperitoneal cavity for the first time in vivo, and the ratio of the dose to the body weight of the giant catfish is 7-12 μg: 1g, phytohemagglutinin (PHA) is injected intraperitoneally for the second time in vivo after 16-20 hours, the ratio of the dose to the body weight of giant barb is 5-10 μg: 1g, and the mass concentration of the injection is 0.1%-0.2% after 4-6 hours of action colchicine, the ratio of the dose to the body weight of the giant catfish is 5-10μg:1g, and the reaction time is 2-4h;

[0022] 2. Sampling: cut the gills of the giant barb treated in step 1, let the blood out for 10-15 minutes, then take out the head kidney an...

specific Embodiment approach 2

[0034] Specific embodiment two: this embodiment is different from collective embodiment one in that: in step two, centrifuge for 5 minutes at a rotational speed of 1000 r / min. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0035] Specific embodiment three: the difference between this embodiment and collective embodiment one or two is: in step four, centrifuge for 5 minutes at a rotation speed of 1000 r / min. Others are the same as in the first or second embodiment.

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Abstract

The invention discloses a method for preparing chromosomes through large-scale barbel fish nephrocyte in-vivo culture and relates to a method for preparing fish chromosomes. The invention aims to solve the technical problem of the method for preparing the chromosomes of the large-scale barbel fish. The method comprises the following steps of: performing living body injection, namely injecting phytohaemagglutinin (PHA) into an abdominal cavity of the living body twice, and injecting colchicine; 2, sampling, namely washing head-kidney in NaCl normal saline, and collecting cells; 3, performing hypotension by using a mixed solution of sodium citrate and KCl; 4, performing cryofixation by using a Carnoy's fixing solution; 5, dropping through a cold drop method; and 6, staining through Giemsa staining solution, and obtaining the chromosomes of the large-scale barbel fish. The method is easy and convenient to operate and stable in result, and the large-scale barbel fish metacinesis specimen which is complete in number, good in dispersion and clear in shape can be obtained. The method is applied to the field of preparation of fish chromosomes.

Description

technical field [0001] The invention relates to a method for preparing fish chromosomes. Background technique [0002] Barbus capito belongs to family Cyprinidae, subfamily Barbinae and genus Barbodes, and is native to the Aral Sea of ​​Uzbekistan. It has excellent characteristics such as wide feeding habits, fast growth, delicious meat, salt-alkali tolerance, and strong adaptability. Its largest individual is 70cm long and 12kg in weight. , translated by Tian Mingcheng. Ichthyology [M]. Beijing: Higher Education Press, 1958: 186-189.). In 2003, the Heilongjiang Fisheries Research Institute introduced this fish into my country, aiming to increase the number of fish cultured in inland saline-alkali waters, so as to promote the development and utilization of saline-alkali waters. [0003] Chromosome is the main carrier of genetic material. Chromosome number and karyotype are one of the main contents of biological cytogenetics research. Genetic breeding provides the basis fo...

Claims

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Application Information

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IPC IPC(8): G01N1/28C12N5/071
Inventor 耿龙武鲁翠云姜海峰徐伟
Owner HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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