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Method for dyeing Giemsa-C band of paris plant cell

A technology of plant cells and dyeing methods, which is applied in the field of Giemsa-C band dyeing of plant cells of the genus Pachyphylla, can solve the problems of unstable dyeing color, difficult analysis, and contamination of film production, and achieve simple and easy-to-operate band patterns, The effect of high resolution and low film contamination

Inactive Publication Date: 2011-09-14
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Hu Fengrong et al. used the improved HBSG method to analyze the Giemsa-C bands on the root tip of Lily lily. During the experimental operation, the temperature and time of hydrochloric acid treatment were not easy to control, which made the bands blurred; The pH of the buffer is easy to change, resulting in unstable dyeing color [Journal of Nanjing Forestry University, Vol. 33, No. 3, pp. 17-19 (2009)]
[0008] Using the above Giemsa-C band staining method for large chromosomes, the research on the Giemsa-C band of the plant of the genus Pachyphylla found that it is prone to problems such as chromosome winding, blurred banding, and serious contamination of the film.

Method used

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  • Method for dyeing Giemsa-C band of paris plant cell
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  • Method for dyeing Giemsa-C band of paris plant cell

Examples

Experimental program
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Effect test

Embodiment 1

[0030] At 10:30, the new white roots of Pachyphysis mosaic plants were taken, and treated in ice-water mixture for 48 hours; fixed with Carnot I for 6 hours, transferred to 70% alcohol, and then stored in a 4°C refrigerator for later use; For a good root tip use 0.2 molL -1 Dissociate with hydrochloric acid at 22°C for 1 hour, wash with distilled water, press with acetic acid with a volume concentration of 45%, and remove the sheet with liquid nitrogen; dehydrate in absolute ethanol for 12 hours, and dry naturally in the room for 2 days; moL -1 Treat in hydrochloric acid for 155s, wash with distilled water; then treat in saturated barium hydroxide solution for 10min, replace and wash with preheated 38°C distilled water until clear, and then wash with 0.05 molL -1 Rinse twice with hydrochloric acid to remove the residual barium carbonate precipitate, and wash the hydrochloric acid with distilled water; after washing, transfer the film to another clean dyeing vat, add 2×SSC, pu...

Embodiment 2

[0033]At 11:30, take the new white root of Huachonglou plant, and treat it in the ice-water mixture for 36 hours; fix it in Carnot Ⅰ for 12 hours, transfer it to 70% alcohol, and store it in the refrigerator at 4°C for later use; use the prepared root tip with 0.2 moL -1 Hydrochloric acid was dissociated at 23°C for 1.5h, washed with distilled water, pressed with acetic acid with a volume concentration of 45%, and removed with liquid nitrogen; dehydrated with absolute ethanol for 12h, and dried naturally in the room for 3d; moL -1 Treat in hydrochloric acid for 155s, wash with distilled water; then treat with saturated barium hydroxide solution for 9min, replace and wash with preheated 38°C distilled water until clarification, and then wash with 0.05 molL -1 Rinse twice with hydrochloric acid to remove the residual barium carbonate precipitate, and wash the hydrochloric acid with distilled water; after washing, transfer the film to another clean dyeing vat, add 2×SSC, put it ...

Embodiment 3

[0036] At 15:30, take the new white roots of Pachyphylla dibaflora and treat them in the ice-water mixture for 48 hours; fix them in Carnot Ⅰ for 6 hours, transfer them to 70% alcohol, and store them in the refrigerator at 4°C for later use; use the prepared root tips with 0.2 molL -1 Dissociate with hydrochloric acid for 1 hour at 24°C, wash with distilled water, press with acetic acid with a volume concentration of 45%, and remove the sheet with liquid nitrogen; dehydrate with absolute ethanol for 12 hours, and dry naturally in the room for 2 days; -1 Treat in hydrochloric acid for 155s, wash with distilled water; then treat in saturated barium hydroxide solution for 10min, replace and wash with preheated 38°C distilled water until clarification, and then use 0.05 molL -1 Rinse twice with hydrochloric acid to remove the residual barium carbonate precipitate, and then rinse the hydrochloric acid with distilled water; after washing, transfer the sheet to another clean dye vat....

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Abstract

The invention belongs to the technical field of cell biology, and in particular relates to a method for dyeing a Giemsa-C band of a paris plant cell. The method comprises the following steps of: (1) preparing materials, namely putting a white young tender root tip of a eugonic paris plant into an ice water mixture for pretreatment, and fixing by using Carnot I for later use; (2) making tablets, namely dissociating the prepared root tip by using hydrochloric acid, tabletting by using acetic acid, peeling by using liquid nitrogen, dehydrating by using absolute ethanol, and naturally drying in aroom; (3) denaturating, namely pre-denaturating by using hydrochloric acid, and denaturating by using barium hydroxide; (4) performing renaturation, namely performing 2*SSC renaturation; (5) dyeing, namely dyeing by using Giemsa dye liquor; and (6) performing microscopic examination by using an optical microscope. The method for dyeing the Giemsa-C band of the paris plant cell is low in tabletting pollution, high in resolution of a banding pattern and stable and simple in the banding pattern and is easy to operate.

Description

technical field [0001] The invention belongs to the technical field of cell biology, in particular to a Giemsa-C band staining method for plant cells of the genus Pachyphylla. Background technique [0002] Chonglou ( Paris ) is Melanthiaceae (Parideae), which is a traditional Chinese herbal medicine in my country. Bite, bruises, convulsions and convulsions embolism. In recent years, it has been widely used in the production of Chinese patent medicines and new medicines. With the development of industrialized production of medicines, the demand for plants of this genus is large, but the growth cycle of its medicinal parts is long, the natural reproduction rate is low, and the growth is slow, which leads to serious excessive digging among the people, and the wild resources are facing depletion, especially some that have not been studied in depth. species are on the verge of extinction. Therefore, it is imminent to study the sustainable utilization of Pachyphysis plant resou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 丁春邦岳海霞李燕周莉君袁明刘静周永红杨瑞武张利
Owner SICHUAN AGRI UNIV
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