A kind of preparation method of the metaphase mitotic phase specimen of plant cell of the genus Neotrichum

A technology for preparing plant cells and specimens, which is applied in the field of preparation of metaphase mitosis specimens of plant cells of the genus Pseudomonas. small damage effect

Inactive Publication Date: 2016-02-10
CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The above-mentioned chemical drugs can prevent or destroy the formation of the spindle body, and are highly toxic and harmful to the body of the experimenter.
The low temperature method takes a long time for pretreatment, and it takes 1 to 3 days for one pretreatment, and the effect is not ideal.

Method used

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  • A kind of preparation method of the metaphase mitotic phase specimen of plant cell of the genus Neotrichum
  • A kind of preparation method of the metaphase mitotic phase specimen of plant cell of the genus Neotrichum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 A method for preparing a chromosomal metaphase sample of a plant of the genus Neotrichum, comprising the following steps:

[0022] ⑴Materials: At 9:00 am, take the vigorously growing potted new wheatgrass in the United States ( P. juncea ), and put it into an open 1.5mL centrifuge tube with water attached to the inner wall.

[0023] (2) At 20°C, place the centrifuge tube with the apex in step (1) in a high-pressure container device, and first inject 0.4MPaN 2 O treatment for 1.5h, and then at the same temperature, N 2 O to make the partial pressure reach 0.75MPa, continue to treat for 2h50min, and obtain the new root after pretreatment.

[0024] (3) Dry the pretreated new roots with filter paper, fix them with Carnot's solution for 20 hours, and obtain the fixed new roots.

[0025] Among them: Carnot's solution is made by mixing absolute ethanol and glacial acetic acid according to the ratio of 3mL: 1mL.

[0026] (4) Rinse the fixed new roots with disti...

Embodiment 2

[0029] Example 2 A method for preparing a chromosomal metaphase sample of a plant of the genus Neotrichum, comprising the following steps:

[0030] ⑴Materials: At 3:00 in the afternoon, the vigorously growing potted new wheatgrass in Kazakhstan ( P. juncea ), and put it into an open 2.0mL centrifuge tube with water attached to the inner wall.

[0031] (2) Under the condition of 25°C, place the centrifuge tube with the apex in step (1) in the high-pressure container device, and first pass 0.4MPaN 2 O treatment for 1.5h, and then at the same temperature, N 2 O to make the partial pressure reach 0.8MPa, continue to treat for 2.5h, and obtain the new root after pretreatment.

[0032] (3) Dry the pretreated new roots with filter paper, fix them with Carnot's solution for 23 hours, and obtain the fixed new roots.

[0033] Among them: Carnot's solution is the same as Example 1 .

[0034] (4) Rinse the fixed new roots with distilled water for 3 times, absorb the water and dis...

Embodiment 3

[0037] Example 3 A method for preparing a chromosomal metaphase sample of a plant of the genus Neotrichum, comprising the following steps:

[0038] ⑴Materials: At 3:00 p.m., vigorously growing potted new wheatgrass in the United States ( P. juncea ), and put it into an open 2.0mL centrifuge tube with water attached to the inner wall.

[0039] (2) At 26°C, place the centrifuge tube with the apex in step (1) in a high-pressure container device, and first inject 0.4MPaN 2 O treatment for 1.5h, and then at the same temperature, N 2 O to make the partial pressure reach 0.8MPa, continue to treat for 2.5h, and obtain the new root after pretreatment.

[0040] (3) Dry the pretreated new roots with filter paper, fix them with Carnot's solution for 23 hours, and obtain the fixed new roots.

[0041] Among them: Carnot's solution is the same as Example 1 .

[0042] (4) Rinse the fixed new roots with distilled water for 3 times, absorb the water and dissociate in glacial acetic acid ...

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Abstract

The present invention relates to a method for preparing a metaphase mitotic phase specimen of plant cells of the genus Pseudotium, which comprises the following steps: (1) taking materials: taking vigorously growing potted new roots of Pseudomonas at any time of the day and putting them into a centrifuge tube; (2) Place the centrifuge tube with the root tip in a high-pressure container, and pretreat it with N2O to obtain the pretreated new root; (3) absorb the water from the pretreated new root, fix it with Carnot’s solution, and obtain the fixed root new roots; (4) the fixed new roots are rinsed and dried and then dissociated with glacial acetic acid to obtain new roots after dissociation; (6) Place the pressed sheet under an optical microscope for observation, select cells with dispersed and clear chromosomes to take pictures for fluorescence in situ hybridization, and finally use a glass knife to mark the position of the cover slip, and put the prepared sheet into Store in a -80°C refrigerator. The invention is efficient and stable.

Description

technical field [0001] The invention relates to a method for preparing a plant chromosome metaphase specimen in the field of cytogenetics, in particular to a method for preparing a plant cell chromosome metaphase specimen of the genus Neotrichum. Background technique [0002] Chromosome preparation is the basic method of cytogenetics research, and it is an indispensable and important means to study biological genetic variation, species evolution, classification, distant hybridization and the relationship between chromosome structure, shape and function. [0003] Chromosome specimen preparation, that is, chromosome preparation, takes the metaphase chromosomes of somatic cell division as the research object, but the duration of metaphase is very short, generally only 10-30 minutes, so that the frequency of metaphase chromosome cells is not high. In order to overcome the above-mentioned contradictions, chemical or physical methods are commonly used to pretreat the material. Whe...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
CPCG01N1/30
Inventor 蔡振媛李媛张毓喻凤刘瑞娟林丽
Owner CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST
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