Preparation method of chromosomes of adult epinephelus akaara

A red-spotted grouper and chromosome technology, applied in the field of cell biology, can solve the problems of long time, difficult acquisition and high cost, and achieve the effect of saving time and increasing the injection concentration

Inactive Publication Date: 2015-04-01
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI +2
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  • Summary
  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, due to the tissue characteristics of the red-spotted grouper, according to the conventional chromosome preparat...

Method used

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  • Preparation method of chromosomes of adult epinephelus akaara
  • Preparation method of chromosomes of adult epinephelus akaara
  • Preparation method of chromosomes of adult epinephelus akaara

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Embodiment Construction

[0027] A method for preparing chromosomes of red-spotted grouper adult fish, which includes pre-treatment of materials, preparation of kidney cell suspension, hypotonic treatment, fixation, dropping and staining, and the specific steps are as follows:

[0028] (1) Pre-processing of materials

[0029] The pre-treatment of the material adopts the in vivo injection method of phytohemagglutinin, and injects PHA to the base of the pectoral fin of the experimental fish at a dose of 15 μg / g fish body weight, and injects colchicine solution at a dose of 2 μg / g fish body weight after 24 hours, about After 3 hours, the tail was docked to lose blood for 20 minutes, and the head kidney was removed;

[0030] (2) Preparation of kidney cell suspension

[0031] Wash the head kidney tissue in 0.8% (weight percent) normal saline to remove blood clots and other tissues, then place it in a petri dish filled with 0.8% (weight percent) normal saline, fully cut it into pieces, and then use four lay...

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Abstract

The invention discloses a preparation method of chromosomes of adult epinephelus akaara and belongs to the field of cell biology. The preparation method comprises the following steps: pretreatment of materials, preparation of kidney cell suspension, low-permeation treatment, fixation, dropping and dyeing, wherein the pretreatment of the materials comprises the steps of with an in vivo injection method of phytohemagglutinin (PHA), injecting PHA into the pectoral-fin base part of the experimental adult epinephelus akaara with 15 micrograms in each gram of the adult epinephelus akaara; after 24 hours, injecting colchicine solution with 2 micrograms in each gram of the adult epinephelus akaara; and after 2.5-3 hours, taking out the head kidney. The preparation method disclosed by the invention has the advantages that the method for one-time injection of PHA is adopted, simultaneously, the injection concentration is increased, and good effect can be obtained after 24 hours, so that the time is saved; and due to the characteristics of tissue cells of the epinephelus akaara, complete cells are not easily obtained when the low-permeation treatment is carried out in the prior art, so that the prior art is creatively changed and a clear chromosome image is obtained.

Description

technical field [0001] The invention belongs to the field of cell biology, and in particular relates to a method for preparing chromosomes of red-spotted grouper adults. Background technique [0002] Chromosomal specimen preparation has the characteristics of simple operation, low price, and low requirement for experimental site. Steps such as slices have been widely used in many fields of biogenetics. However, due to the tissue characteristics of the red-spotted grouper itself, according to the conventional chromosome preparation method, it is difficult to obtain, the cost is high, the effect of making chromosome samples is poor, and the time is long. Contents of the invention [0003] The technical problem to be solved by the present invention is to provide a method for preparing chromosomes of red-spotted grouper adult fish. The present invention improves the conventional chromosome preparation method, changes the hypotonic treatment time and pretreatment method, and o...

Claims

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Application Information

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IPC IPC(8): G01N1/30
Inventor 陈超刘莉孔祥迪李炎璐李娟徐万土陈建国于欢欢
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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