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Preparation method of turbellarian worm chromosome specimen

A chromosome and planarian technology, applied in the field of cytogenetics, can solve the problems of difficult sampling and processing, unfavorable research and analysis, and unclear production, and achieve the effect of clear background, good cell dispersion, and favorable karyotype analysis

Inactive Publication Date: 2014-08-13
ZHOUKOU NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The existing mature animal chromosome specimen preparation methods are mostly suitable for higher vertebrates and humans, and can be prepared after processing by collecting bone marrow or blood; while planarians only have tissue fluid, no spinal cord and blood, so sampling and processing are difficult
Most of the existing technologies use the method of Li Guangpeng ("Journal of Zoology" 1992, 27 (5), 30~31), but the results are unstable and the film production is not clear, which is not conducive to research and analysis, and there is a large room for improvement

Method used

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  • Preparation method of turbellarian worm chromosome specimen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] (1) Material collection and pretreatment: Select a lively and healthy planarian with a body length of 3 cm, put it in dechlorinated tap water, and starve it for 1 week until no metabolic waste is discharged;

[0030] (2) Regeneration tissue culture: place the planarian treated in step (1) on a glass slide, and when it is fully stretched, cut the planarian into 3 sections with an ethanol-sterilized blade, and place it at a constant temperature of 10°C Cultivate in sterile water in the incubator for 3 days, and white regenerative tissue grows on the section;

[0031] (3) Tissue crushing and colchicine treatment: Carefully remove the regenerated planarian tissue obtained in step (2) with a clean blade, place it in a 1.5mL centrifuge tube, break it slightly with a small scissor needle, and then add 0.2g / L of Colchicine aqueous solution 0.5mL, stand at 10°C for 2.5 hours, then centrifuge at 800rpm for 6 minutes, discard the supernatant;

[0032] (4) KC1 hypotonic treatment:...

Embodiment 2

[0039] (1) Material collection and pretreatment: Select a lively and healthy planarian with a body length of 5 cm, put it in dechlorinated tap water, and starve it for 2 weeks until no metabolic waste is discharged;

[0040] (2) Regeneration tissue culture: place the planarian treated in step (1) on a glass slide, and when it is fully stretched, cut the planarian into 5 sections with an ethanol-sterilized blade and place it at a constant temperature of 15°C Cultivate in sterile water in the incubator for 5 days, and white regenerative tissue grows on the section;

[0041] (3) Tissue crushing and colchicine treatment: Carefully remove the regenerated planarian tissue obtained in step (2) with a clean blade, place it in a 2.0mL centrifuge tube, break it slightly with a small scissor needle, and then add 0.25g / L of Colchicine aqueous solution 1.0mL, let stand at 15°C for 3.5 hours, then centrifuge at 1000rpm for 5 minutes, discard the supernatant;

[0042] (4) KC1 hypotonic trea...

Embodiment 3

[0049] (1) Material collection and pretreatment: Select a lively and healthy planarian with a body length of 4 cm, put it in dechlorinated tap water, and starve it for 10 days until no metabolic waste is discharged;

[0050] (2) Regenerative tissue culture: place the planarian treated in step (1) on a glass slide, and when it is fully stretched, cut the planarian into 4 sections with an ethanol-sterilized blade and place it at a constant temperature of 12°C Cultivate in sterile water in the incubator for 4 days, and white regenerative tissue grows on the section;

[0051] (3) Tissue crushing and colchicine treatment: Carefully remove the regenerated planarian tissue obtained in step (2) with a clean blade, place it in a 1.5mL centrifuge tube, break it slightly with a small scissor needle, and then add 0.2g / L of Colchicine aqueous solution 0.5mL, stand at 15°C for 3 hours, then centrifuge at 900rpm for 5 minutes, discard the supernatant;

[0052] (4) KC1 hypotonic treatment: a...

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Abstract

The invention relates to a preparation method of a turbellarian worm chromosome specimen. The preparation method of the turbellarian worm chromosome specimen comprises the steps of material preparation, pretreatment, regenerated tissue culture, tissue smashing, centrifugal suspending, colchicine blockage, KCl low permeability treatment, centrifugal suspending and dripping, dyeing, baking and sheet sealing. By adopting the preparation method, regenerated tissues and metaphase cells are fixed and treated fully, the chromosome preparation background is clear, cell dispersion is high, good cell scattering is realized, the number of metakinesis phases is large, and karyotype analysis is facilitated.

Description

technical field [0001] The invention belongs to the field of cytogenetics, and in particular relates to a method for preparing a planarian chromosome sample. Background technique [0002] Planarians are widely distributed in my country. In the history of animal evolution, planarians are the first group with bilateral symmetry and three germ layers. Their evolutionary status is very important, and they are good materials for the research of developmental biology, cytogenetics and regenerative biology. Chromosome preparation of planarian is the basis of its genetics research, as well as the basis of cell and molecular biology research. [0003] The existing mature methods for preparing animal chromosome specimens are mostly suitable for higher vertebrates and humans, and can be prepared after processing by collecting bone marrow or blood; while planarians only have interstitial fluid, no spinal cord and blood, so sampling and processing are difficult. Li Guangpeng (Journal of...

Claims

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Application Information

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IPC IPC(8): G01N1/30
Inventor 马克世盛东峰武安泉王永立杨同文陈龙
Owner ZHOUKOU NORMAL UNIV
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