Fish embryo chromosome flaking method

A chromosome production and fish technology, applied in the field of genetic engineering, can solve the problems affecting the stability of production results, time-consuming and laborious operation, experimental failure, etc., and achieve the effects of shortening sampling time, simple and efficient operation, and shortening sampling time.

Inactive Publication Date: 2010-09-08
YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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Problems solved by technology

The advantage of the single-embryo slice method is that it can accurately identify the ploidy of a single embryo. The disadvantage is that the operation is delicate, time-consuming and labor-intensive. When the number of embryos to be identified is large, the application of this method is limited. For example, when calculating the induction rate, at least More than 30 embryos are needed, and the traditional method of mixed embryo production can solve t

Method used

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Embodiment

[0018] Utensils and reagents: 5ml conical graduated centrifuge tube, pipette, wide-mouth pipette, tweezers, frozen slides, 10ml beaker, RPMI 1640 cell culture medium, physiological saline for fish: 0.75% sodium chloride solution, colchicine solution 100 μg / Milliliter, hypotonic solution: 0.0375mol / L potassium chloride solution, fixative: methanol and glacial acetic acid mixed at a volume ratio of 3:1, Giemsa staining mother solution.

[0019] Its operation method is as follows:

[0020] (1) Sampling: get 100 yellow catfish eggs, put them into a 10ml beaker, wash them twice with 0.75% fish saline, blow and beat them with a straw to remove surface dirt;

[0021] (2) Cell culture and collection: After the sample is cleaned, pour out the fish with normal saline, then add 4ml of 1640 cell culture medium containing 10 μg / ml colchicine, and treat for 60 minutes. Clamp the eggs together to free the embryonic cells. After all the fish eggs are clamped, use a large-mouth straw to blow...

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Abstract

The invention discloses a fish embryo chromosome flaking method, relating to genetic engineering. The method includes the steps of taking out 100-200 roes during the middle-stage gastrula and the late-stage gastrula, washing the roes for twice through normal saline for fish to remove the membranes and the vitellines of the roes, adding the roes into colchicine-containing 1640 culture solution for cell culture, striking the roes to enable the cells to be even, adding fixative to fix the cells, drying and dyeing through staining mother solution. The invention overcomes the defects that the conventional method is time-consuming and labor-consuming and batch production can not carried out; and the fish embryo chromosome flaking method improves the stability of the flakes and the accuracy of the inductivity. Therefore, the fish embryo chromosome flaking method provides a reliable guarantee for genetic breeding and breed improvement of fish.

Description

technical field [0001] The invention relates to genetic engineering, in particular to a method for making fish embryo chromosome specimens. Background technique [0002] It is a development direction in the fishery to apply new technologies of bioengineering to fish genetic breeding and species improvement, especially fish cell engineering such as polyploid induction, which uses chromosomes as the object of operation, has shown broad application prospects . At present, the methods for ploidy detection of fish embryos mainly include single embryo preparation method and mixed embryo preparation method. The advantage of the single-embryo slice method is that it can accurately identify the ploidy of a single embryo. The disadvantage is that the operation is delicate, time-consuming and labor-intensive. When the number of embryos to be identified is large, the application of this method is limited. For example, when calculating the induction rate, at least More than 30 embryos ...

Claims

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Application Information

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IPC IPC(8): G01N1/30G01N1/02
Inventor 梁宏伟李忠邹桂伟吴勤超罗相忠
Owner YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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