Method for preparing chromosome of procambarus clarkii

A technology for procrayfish and chromosomes, which is applied in the field of preparation of chromosomes of Crayfish, can solve the problems of difficult chromosome division in metaphase, large number, small chromosomes and the like, and achieves easy observation, high number accuracy and shallow background. Effect

Inactive Publication Date: 2017-09-29
HUAZHONG AGRICULTURAL UNIVERSITY
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Problems solved by technology

[0004] The object of the present invention is to provide a kind of Procambarus clarkii with good chromosome dispersion, recognizable and countable morphology, aiming at the problems that Procambarus clarkii chromosomes are small, large in number, difficult to obtain dispersed chromosome division metaphase, and sample collection time is limited. Improved method for preparing Procambarus clarkii chromosomes, the sample collection time involved in the present invention is not limited by seasons

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  • Method for preparing chromosome of procambarus clarkii
  • Method for preparing chromosome of procambarus clarkii

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Embodiment 1

[0025] The preparation of the chromosome of embodiment 1 Procambarus clarkii adult shrimp

[0026] 1. Preparation of reagents and related materials:

[0027] (1) Preparation of normal saline:

[0028] 7.5g NaCl;

[0029] 0.2g CaCl2;

[0030] 0.2g KCl;

[0031] 0.02g NaHCO2;

[0032] Dilute to 1000 mL with deionized water.

[0033] (2) Phytohemagglutinin (PHA): the concentration is 2 mg / mL, and the solvent is sterilized physiological saline.

[0034] (3) Colchicine: 1 mg / mL, the solvent is sterilized physiological saline.

[0035] (4) 0.75% KCl solution: Dissolve 0.75g of KCl in 100mL sterilized ddH 2 O middle;

[0036] (5) Fixative-1: Prepared according to the volume ratio of methanol and glacial acetic acid at 3:1, prepared immediately for use, and pre-cooled on ice.

[0037] (6) Fixative-2: Prepared according to the volume ratio of methanol and glacial acetic acid at 1:1, prepared immediately for use, and pre-cooled on ice.

[0038] (7) Giemsa staining solution: Wh...

Embodiment 2

[0048] Chromosome preparation of embodiment 2 Procambarus clarkii juvenile shrimp

[0049] Choose young Procambarus clarkii shrimp that are healthy, harmless and vigorous, with a weight of about 15-20g. The preparation of experimental reagents and related materials and the experimental steps are the same as in Example 1, and the results of microscopic examination are shown in figure 2 .

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Abstract

The invention belongs to the technical field of zoocytologic experiments and particularly relates to a method for preparing a chromosome of procambarus clarkii. The method comprises the steps: separately carrying out injection treatment on procambarus clarkii bodies by using phytohemagglutinin PHA and colchicine, then, taking antennal glands of the procambarus clarkii, rinsing the antennal glands in normal saline, then, shearing tissue into muddy tissue, and carrying out a centrifugation method, so as to obtain a cell suspension; separately carrying out hypotonic treatment and pre-immobilization by using a 0.75% KCl solution and an immobilization solution-1; then, carrying out immobilization on cells separately by using the immobilization solution-1 and an immobilization solution-2 which are in the ratio of 3:1; obliquely placing a precooled glass slide, sucking a proper volume of cell suspension by a pipette, vertically spraying the cell suspension onto the glass slide in a rapid and powerful manner, rapidly baking the glass slide over flame of an alcohol lamp for 2 to 3 seconds, and airing the glass slide in; staining the glass slide accompanied by the chromosome with a Giemsa staining solution; carrying out microscopical observation and photographing. The method has the advantages of relatively good chromosome dispersion, discernible form and countable chromosome split phase.

Description

technical field [0001] The invention relates to the technical field of animal cell experiments, in particular to a method for preparing the chromosome of Procambarus clarkii. Background technique [0002] Procambarus clarkii (Procambarus clarkii), belonging to Crustacea, Decapod order, Crawfish family, Procambarus genus, is native to northern Mexico and southern United States. Introduced into my country around 1929, the shrimp has become an important aquaculture animal in China and has made certain contributions to my country's economy. Understanding the karyotype characteristics of this shrimp can lay a certain theoretical foundation for further research on the genetic breeding, variety improvement and control of this shrimp. [0003] Because the chromosomes of Procambarus clarkii are small in shape and large in number, it is difficult to obtain well-dispersed and clearly identifiable chromosome pictures by general chromosome preparation methods, which brings certain diffi...

Claims

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Application Information

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IPC IPC(8): G01N1/28
CPCG01N1/28
Inventor 李艳和石林林张龙周小云王卫民
Owner HUAZHONG AGRICULTURAL UNIVERSITY
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