Microdissection technology of tobacco chromosomes

A technique of microdissection and chromosome, which is applied in the biological field to achieve the effect of multiple cleavage phases, simple process and high degree of dispersion

Inactive Publication Date: 2017-05-10
GUIZHOU NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0010] The technical problem to be solved by the present invention is to establish an efficient and accurate laser microdissection technology for tobacco that is easy to operate, completely recovers the target chromosome and is pollution-free, based on the technology of chromosome removal and low-osmosis method

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  • Microdissection technology of tobacco chromosomes
  • Microdissection technology of tobacco chromosomes

Examples

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Embodiment 1

[0025](1) Chromosomal specimen preparation: Sow the mature and plump tobacco seeds in distilled water at 40°C for 50 minutes, sow them on damp filter paper, place them in a petri dish (put a piece of filter paper on the bottom of the petri dish to keep it moist), and germinate at 26°C . When the root grows to 0.5-1 cm, cut the fresh root tip between 9:00-11:00 in the morning, and put it into saturated α-bromine solution for pretreatment at room temperature for 5 hours. Wash with distilled water three times, and fix overnight with Carnot's fixative solution (ethanol: glacial acetic acid = 3:1). After washing with distilled water for 3 times, hypotonic for 1 h before placing in distilled water, treat with 3% cellulose-pectinase mixed enzyme solution at 37 °C for 2.5 h, wash 3 times, and hypotonic for 60 min after placing in distilled water. Cut out 1-2mm root apical meristem tissue on a clean glass slide, drop 1 drop of fixative solution, quickly crush the material with tweezer...

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Abstract

The invention discloses a microdissection technology of tobacco chromosomes. The microdissection technology is characterized by comprising the following steps: (1) preparing a chromosome specimen: after immersing ripe and full seeds of tobaccos, putting the seeds into a culture dish and germinating; after roots grow to 0.5cm to 1cm, scissoring fresh root tips from 9:00 to 11:00 in the morning; putting the roots into a saturated alpha-bromnaphthalene solution and pre-treating at room temperature; washing and fixing by Carnoy I stationary liquid for one night; washing and putting the roots into distilled water and carrying out low percolation; treating with cellulose-pectinase mixed enzyme liquid and washing; putting the roots into the distilled water and carrying out the low percolation; putting root tip meristems on a slide and dropwise adding the stationary liquid; crushing materials and coating, and removing large tissue residues; then adding the stationary liquid; dyeing and washing to remove a dyeing solution; marking a good metaphase of microscope inspection for later use; (2) carrying out microdissection on the chromosomes: putting the specimen slide onto an objective table of a laser microdissection system and setting a cutting path; mounting a centrifugal tube with a viscous tube cover on an automatic recycling device of the laser microdissection system; treating the viscous centrifugal tube for later use after the chromosomes are cut and recycled.

Description

technical field [0001] The invention relates to a tobacco chromosome microdissection technology, which belongs to the field of biotechnology. Background technique [0002] Plant root tip cell mitosis and chromosome production technology: The meristematic tissues of vigorous growth of plants, such as root tip, shoot tip, and young leaves, are undergoing mitosis. Take these tissues as materials, kill the cells quickly after fixing, and keep the substances and living conditions in the cells, and then go through dissociation, staining, pressing and other processes to quickly disperse and attach the cells to the glass slide and Between the coverslips, a large number of cells in various stages of mitosis can be seen under the microscope, and the chromosomes can be observed. In metaphase, the centromeres of each chromosome are arranged on the equatorial plane, and the two arms extend to both sides of the equatorial plane. The spindle is fully formed, with one end attached to the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28
CPCG01N1/286G01N2001/2886
Inventor 盛茂银任学良王霖娇
Owner GUIZHOU NORMAL UNIVERSITY
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