A method for fluorescent in situ hybridization of Mongolian leek chromosome

A fluorescence in situ hybridization and chromosome technology, applied in the field of cytogenetics and molecular biology, to achieve the effect of strong hybridization specificity, time saving and high hybridization specificity

Active Publication Date: 2020-06-09
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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  • A method for fluorescent in situ hybridization of Mongolian leek chromosome
  • A method for fluorescent in situ hybridization of Mongolian leek chromosome
  • A method for fluorescent in situ hybridization of Mongolian leek chromosome

Examples

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Embodiment 1

[0045] This embodiment provides a method for fluorescence in situ hybridization of chromosomes of Allium mongolica, which includes the following steps:

[0046] S1. Preparation of chromosome slide specimens of Allium mongolicum:

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

[0048] (2) Tablet preparation: Put the cut root tips into a vial, add appropriate amount of water, cover, and 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 pretreated root tips and fix them with freshly prepared Carnot's fixative (glacial acetic acid: alcohol=1:3) at room temperature (25°C) for 12-24 hours.

[0050] (4) Dissociation: Take the ...

Embodiment 2

[0068] This embodiment provides a method for fluorescence in situ hybridization of chromosomes of Allium mongolica, which includes the following steps:

[0069] S1. Preparation of chromosome slide specimens of Allium mongolicum:

[0070] (1) Material: Take Mongolian leek seeds and rinse them with distilled water for 1-3 times, then soak them in the seed soaking solution for 0.5 hours, and then place them in a petri dish with 2 layers of wet filter paper, add water and place in a thermostat at 22- Cultivate for 18 hours at 25°C, the root tip of the seed grows to 0.8cm, which meets the requirements of the seed root tip in the range of 0.5-1cm, and then cut the root tip 1-2mm; the soaking liquid contains 80mg / L brown sugar, 150mg / L mannitol and A mixed aqueous solution of 25mg / L acetone.

[0071] (2) Tablet preparation: Put the cut root tips into a vial, add appropriate amount of water, cover, and quickly place it in a mixture of ice and water at 0°C, and pretreat it in a refrigerator ...

Embodiment 3

[0091] This embodiment provides a method for fluorescence in situ hybridization of chromosomes of Allium mongolica, which includes the following steps:

[0092] S1. Preparation of chromosome slide specimens of Allium mongolicum:

[0093] (1) Material: Take Mongolian leek seeds and rinse 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 thermostat at 22- Cultivate for 21 hours at 25°C, the root tip of the seed grows to 0.6cm, which meets the requirements of the seed root tip of 0.5-1cm, and then cut the root tip 1-2mm; the seed soaking solution contains 90mg / L brown sugar, 120mg / L mannitol and A mixed aqueous solution of 40mg / L acetone.

[0094] (2) Tablet preparation: Put the cut root tips into a vial, add appropriate amount of water, cover, and quickly place it in a mixture of ice and water at 0°C, and pretreat it in a refrigerator at 0...

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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 specifically relates to a method for fluorescence in situ hybridization of Mongolian leek chromosomes. Background technique [0002] Fluorescence in situ hybridization (FISH) is an important non-radioactive labelled in situ hybridization technology developed in the late 1980s. It is used in the research of plant molecular cytogenetics, gene amplification, gene mapping and Plant evolution and identification of genetic relationships have been widely used. 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 genes are one of the most important housekeeping genes in organisms, and their transcription products are assembled together with ribosomal proteins to form riboso...

Claims

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Application Information

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