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FISH (fluorescence in situ hybridization) method of allium mongolicum regel chromosome

A fluorescence in situ hybridization and chromosome technology, applied in the field of cytogenetics and molecular biology, to achieve the effects of shortened culture time, strong fluorescent signal, and strong hybridization specificity

Active Publication Date: 2019-01-08
上海秋皇医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, the karyotype analysis of Mongolian Chinese chives still adopts classical cytological methods, and there are no related reports on the application of fluorescence in situ hybridization method to Mongolian Chinese chives for karyotype analysis.

Method used

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  • FISH (fluorescence in situ hybridization) method of allium mongolicum regel chromosome
  • FISH (fluorescence in situ hybridization) method of allium mongolicum regel chromosome
  • FISH (fluorescence in situ hybridization) method of allium mongolicum regel chromosome

Examples

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Embodiment 1

[0045] This embodiment provides a method for fluorescence in situ hybridization of the Mongolian Chinese chives chromosome, comprising the following steps:

[0046] S1. Preparation of Mongolian leek chromosome slide specimen:

[0047] (1) Material collection: Take Mongolian leek seeds and place them in a petri dish with 2 layers of wet filter paper, add water and place them in an incubator at 22-25°C for 4-5 days, and the root tips of the seeds will grow to 0.6cm, which meets the requirements of The required range of the root tip of the seed is 0.5-1cm, and then cut the root tip of 1-2mm.

[0048] (2) Tablet preparation: put the excised root tip into a vial, add an appropriate amount of water, seal the cap, quickly place it in a mixture of ice and water at 0°C, and pretreat it in a refrigerator at 0°C for 20-24 hours.

[0049] (3) Fixation: Rinse the root tip after the above pretreatment, and fix it at room temperature (25° C.) for 12-24 hours with a freshly prepared Carnoy’s...

Embodiment 2

[0068] This embodiment provides a method for fluorescence in situ hybridization of the Mongolian Chinese chives chromosome, comprising the following steps:

[0069] S1. Preparation of Mongolian leek chromosome slide specimen:

[0070] (1) Material collection: Take Mongolian leek seeds and wash them with distilled water for 1-3 times, then soak them in the seed soaking solution for 0.5 hours, then place them in a petri dish with 2 layers of wet filter paper, add water and place them in a constant temperature box at 22- Cultivate at 25°C for 18 hours, the root tip of the seed grows to 0.8cm, which meets the requirements of the root tip of the seed in the range of 0.5-1cm, and then cuts the root tip of 1-2mm; Mixed aqueous solution of 25mg / L acetone.

[0071] (2) Tablet preparation: put the excised root tip into a vial, add an appropriate amount of water, seal the cap, quickly place it in a mixture of ice and water at 0°C, and pretreat it in a refrigerator at 0°C for 20-24 hours...

Embodiment 3

[0091] This embodiment provides a method for fluorescence in situ hybridization of the Mongolian Chinese chives chromosome, comprising the following steps:

[0092] S1. Preparation of Mongolian leek chromosome slide specimen:

[0093] (1) Material collection: Take Mongolian leek seeds and wash them with distilled water for 1-3 times, then soak them in the seed soaking solution for 0.5 hours, then place them in a petri dish with 2 layers of wet filter paper, add water and place them in a constant temperature box at 22- Cultivate at 25°C for 21 hours, the root tip of the seed grows to 0.6cm, which meets the requirements of the root tip of the seed in the range of 0.5-1cm, and then cuts the root tip of 1-2mm; Mixed aqueous solution of 40mg / L acetone.

[0094] (2) Tablet preparation: put the excised root tip into a vial, add an appropriate amount of water, seal the cap, quickly place it in a mixture of ice and water at 0°C, and pretreat it in a refrigerator at 0°C for 20-24 hours...

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Abstract

The invention provides a FISH (fluorescence in situ hybridization) method of an allium mongolicum regel chromosome. The method comprises the steps as follows: S1, preparation of a allium mongolicum regel chromosome slide specimen; S2, preparation o a 45S rDNA probe: selecting a section of conserved sequence from a 45S rDNA sequence of corn to synthesize the probe, and linking a fluorophore at the5' end or 3' end of the probe sequence, wherein the probe sequence is any one nucleotide sequence shown in SEQ ID NO:1-3; S3, FISH; S4, elution and detection. By means of the FISH method, fish can beperformed on the allium mongolicum regel, the 45S rDNA site of the allium mongolicum regel chromosome can be clearly displayed, the hybridization specificity is high, and the method can be better applied to detection of the allium mongolicum regel chromosome and facilitate analysis of the karyotype of the allium mongolicum regel chromosome.

Description

technical field [0001] The invention belongs to the technical field of cytogenetics and molecular biology, and in particular relates to a fluorescent in situ hybridization method of the Mongolian allium chromosome. Background technique [0002] Fluorescence in situ hybridization (FISH) is an important non-radioactive labeling in situ hybridization technique developed in the late 1980s. It is used in the study of plant molecular cytogenetics, gene amplification, gene mapping and It has been widely used in the identification of plant evolution and kinship. Repeated DNA sequences and multi-gene families are used as probes to hybridize with chromosomes in situ, which is easy to produce higher resolution, because these DNA multi-copy sequences can reach dozens of chromosomes. kb or several Mb. [0003] Ribosomal rRNA gene is one of the most important housekeeping genes in organisms, and its transcripts are assembled into ribosomes together with ribosomal proteins. The main rDNA...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6841C12Q1/6806
CPCC12Q1/6806C12Q1/6841C12Q2563/107
Inventor 张边江陈全战唐宁王立科杨平钱保俐
Owner 上海秋皇医药科技有限公司
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