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Method for preparing Leuciscus sp. chromosomes

The technology of yalong fish and chromosome is applied in the field of fish chromosome preparation, which can solve the problems of long cell culture process, difficult blood collection from fish and high culture conditions, and achieve the effects of short cell culture time, easy culture and high culture conditions.

Inactive Publication Date: 2018-12-18
HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a method for preparing the chromosome of Arowana in order to solve the technical problems that it is not easy to collect blood from fish that are too small in the blood cell culture method, the cell culture process is long, the culture conditions are high, and the culture is difficult.

Method used

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  • Method for preparing Leuciscus sp. chromosomes
  • Method for preparing Leuciscus sp. chromosomes

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specific Embodiment approach 1

[0017] Specific implementation mode one: a method for preparing a dragon fish chromosome in this embodiment is carried out according to the following steps:

[0018] 1. Breed 10-month-old Yalong fish with a body length of 10.20-15.17cm, a weight of 13.20-37.56g, and 10 months old in an inflatable aquarium with a water temperature of 15°C±1°C for 1 week;

[0019] 2. According to the ratio of PHA and fish body weight of 10μg:1g, inject PHA to Yalong fish. After 16-18h, inject colchicine solution according to the ratio of colchicine solution and fish body weight of 1-2μg:g, after 2-4h Cut off the gills and cut off the tail, let the blood out in the water, dissect the fish body, take out the head kidney, put it in a plate filled with 0.75% normal saline for cleaning, remove the blood clots, put it in a petri dish filled with normal saline, and use tweezers Grind the head kidney tissue (to free the cells in it), inhale into a 10mL centrifuge tube and pipette repeatedly, add normal ...

specific Embodiment approach 2

[0024] Specific embodiment two: the difference between this embodiment and specific embodiment one is that in step 2, according to the ratio of PHA and fish body weight 10 μ g: 1 g, the Yalong fish is injected with PHA, and after 17 hours, the ratio of colchicine solution and fish body weight is 1 μ g: g The ratio of colchicine solution was injected again. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0025] Specific embodiment three: the difference between this embodiment and specific embodiment one or two is that after 3 hours in step two, the gills are cut off and the tail is cut off to let blood in water, the fish body is dissected, the head kidney is taken out, and placed in a physiological tank with a mass concentration of 0.75%. Wash in a flat dish with saline, remove blood clots, place in a petri dish filled with normal saline, grind the head kidney tissue with tweezers to obtain a cell suspension. Others are the same as in the first or second embodiment.

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Abstract

The invention discloses a method for preparing Leuciscus sp. chromosomes, and relates to a method for preparing fish chromosomes. The invention aims to solve the technical problems that the blood cells culturing method is difficult to collect blood, long in cells culturing process, high in culturing condition and huge in culturing difficulty for undersized fishes. The method comprises the following steps: 1) injecting PHA and colchicine, preparing a cell suspension, centrifuging, adding a hypotonic solution and centrifuging, and discarding supernatant to obtain a cell pellet; 2) fixing; 3) adding 1 to 1.2ml Carnoy's fixative to the centrifuged cell pellet, uniformly mixing to prepare a cell suspension, sucking the cell suspension into a rubber head dropper, adding 2 to 3 drop of the cell suspension under conditions that the rubber head dropper is 25 to 30cm away from a frozen slide and the angle between the rubber head dropper and the frozen slide is 35 to 50 degrees, uniformly bakingthe frozen slide over flames of an alcohol lamp for 3 to 5 times, and drying naturally at room temperature to obtain the Leuciscus sp. chromosomes. The method according to the invention is short in cells culturing time and easy to culture. The Leuciscus sp. chromosomes obtained by the method according to the invention have more spitting phases and have a good effect.

Description

technical field [0001] The invention relates to a method for preparing fish chromosomes. Background technique [0002] Leuciscus sp. is a hybrid fish (Hybrid dace) produced by the interspecific crossbreeding of Leuciscus waleckii Dybowski × Leuciscus idus Linnaeus in Dali Lake. The trade name is Yalong fish; Dari The lake crocodile and the high-body crocodile belong to the order Cypriniformes, Cyprinidae, and the genus Leuciscus of the subfamily Leuciscinae. The Heilongjiang Fisheries Research Institute of the Chinese Academy of Fishery Sciences, with the support of the Heilongjiang Provincial Science and Technology Research Project (Project No.: GCI2B307), completed the early stage creation of Yalong fish germplasm, determination of growth traits, pond cultivation and production selection. Practice has proved that the Yalong fish, a hybrid species of Washii Yaluo fish and Gaoti Yaluo fish, has more growth advantages than its parents, and is deeply loved by the majority of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G01N1/30G01N1/38
CPCG01N1/2813G01N1/286G01N1/30G01N1/38G01N2001/2866G01N2001/305G01N2001/386
Inventor 苏宝锋梁利群薛淑群尚梅孙博常玉梅
Owner HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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