Preparation method of siganus guttatus chromosome

A technology for spotting basketfish and chromosomes, which is applied in the field of preparation of basketfish chromosomes, which can solve the problems of inconspicuous difference in chromosome shape and large number of chromosomes, and achieve the effect of moderate length, clear centromere and complete number

Inactive Publication Date: 2012-11-07
EAST CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Abstract
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  • Application Information

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Problems solved by technology

[0004] The technical problem to be solved by the present invention is that the number of chromosomes of the existing seawater fish is large and small, and the difference in the chromosome shape of the same cell in the middle stage of division is often not very obvious. The preparation method can provide a reliable technical means for exploring the cytogenetic characteristics of the fish, the protection of germplasm resources, the evolution and classification of the system, and the development of cross-breeding experiments

Method used

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  • Preparation method of siganus guttatus chromosome

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Embodiment 1

[0024] 1) Take out the head-kidney tissue from the fish, put it in a petri dish filled with 0.9wt% normal saline, wash it 2-3 times, remove blood clots and other tissues, and cut the head-kidney tissue with scissors.

[0025] 2) Use a pipette to transfer the fully chopped head kidney tissue into a 10ml glass homogenizer, and fully homogenize to make a cell suspension.

[0026] 3) Use a pipette to transfer the cell suspension into a 10ml centrifuge tube, add 0.075mol / L KCl solution, put it in a 37℃ constant temperature water bath for 30min, and centrifuge at 1000r / min for 8min.

[0027] 4) Collect the cells after hypotonicity, add Carnoy’s fixative (pre-cooled to 4°C) (methanol: glacial acetic acid = 3:1, ready-to-use), pipette evenly, let stand at 4°C for 30min, 1000r / min centrifuged for 8min, centrifuged and then fixed once.

[0028] 5) After fixation and centrifugation, discard the supernatant, add pre-cooled to 4°C fixative solution (methanol: glacial acetic acid = 1:1, r...

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Abstract

The invention relates to a preparation method of a siganus guttatus chromosome. The preparation method comprises the following steps of: (1) taking out head-kidney tissues from siganus guttatus, placing in a culture dish containing physiological saline, cleaning, removing blood cots and other tissues, cutting the head-kidney tissues into pieces, and then fully homogenizing the head-kidney tissues to prepare a cell suspension; (2) performing hypotonicity and fixation; (3) performing fixed centrifugation, and then drilling on a slide; and (4) dyeing, drying the slide, then placing the slide under a microscope for observation, and selecting metaphase cells with good dispersion of the chromosome for performing photomicrometrology. The preparation method disclosed by the invention is simple and convenient in operation and stable in result, expensive reagents and special instruments are not required, and the metaphase mitosis in the siganus guttatus chromosome, which has the advantages of complete number, good dispersion, appropriate length, clear centromeres, appropriate separation of two monomers and clear shape, can be obtained by making the slide.

Description

technical field [0001] The invention belongs to the field of preparation of fish chromosomes, and in particular relates to a method for preparing chromosomes of fish-spotted fish. Background technique [0002] Basketfish (Siganus guttatus), belonging to the class Osteichthyes, subclass Actinopterygii, order Perciformes, suborder Acanthuroidei, family Siganidae, basket fish The genus is an omnivorous, euryhaline, warm-water fish that is distributed from coral reefs to estuary waters. Basket fish is delicious, with high economic and nutritional value. It is popular among consumers in Guangdong, Hong Kong, Southeast Asia, the Middle East, Fiji and other countries and regions. [0003] Chromosomal karyotype analysis is one of the important contents of fish cytogenetics research. It is not only of great significance for understanding the taxonomic system, evolutionary relationship and chromosome evolution process of fish, but also provides a cytogenetic basis for fish genetic br...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 宋炜马凌波张凤英章龙珍张涛蒋科技
Owner EAST CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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