Fluorescence in-situ hybridization method for asparagus fern medium-term chromosomes

A fluorescence in situ hybridization and chromosome technology, applied in the field of molecular cytogenetics, can solve the problems such as the application of fluorescence in situ hybridization technology, the failure of chromosome production, the expansion and deformation of chromosomes, etc., to achieve clear signal points, high labeling efficiency, full effect of denaturation

CN106399499AInactive Publication Date: 2017-02-15XINXIANG MEDICAL UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Publication Date
2017-02-15
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention provides a fluorescence in-situ hybridization method for asparagus fern medium-term chromosomes. An asparagus fern medium-term chromosome specimen is prepared by a drop-spreading technique, and by the method, a high-quality asparagus fern chromosome preparation can be obtained. On the basis, fluorescein is used for marking a 45 S rDNA or / and 5D rDNA probe; finally, fluorescence in-situ hybridization of the chromosomes is carried out; and after hybridization of the chromosomes by the probe, fluorescence signals can be directly detected, and therefore, the copy number and the position of a targeted DNA fragment on the chromosomes are determined. The asparagus fern chromosomes which are obtained by the preparation and the fluorescence in-situ hybridization method are complete in morphology, high in hybridization specificity and good in repeatability, operation steps are simple, a large amount of time can be saved, and the fluorescence in-situ hybridization method can be used for identification and structure research of the asparagus fern chromosomes, and provides a new path for evolution research and the like of asparagus linn.
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Description

technical field

[0001] The invention belongs to the technical field of molecular cytogenetics, and in particular relates to a fluorescence in situ hybridization method for asparagus asparagus metaphase chromosomes. Background technique

[0002] Asparagus asparagus, also known as Yunpian pine, Asparagus thorns, and Yunzhu, is a perennial herbaceous vine of the genus Asparagus in the family Liliaceae. Asparagus asparagus has soft leaves, elegant bamboo shape, and is green all the year round. It is a famous indoor potted flower and an excellent lining material for small bonsai and cut flowers. It has high ornamental value. Its root can be used as medicine, can cure acute bronchitis, and has the function of moistening lung and relieving cough. At present, the research on Asparagus asparagus mainly focuses on cultivation techniques and other aspects, and the research on its cytology is relatively lagging behind. This is mainly because the seeds of Asparagus asparagus are difficu...

Examples

Embodiment

[0025] (1) Preparation of chromosomal slide specimens. Control the water of potted asparagus bamboo for three days, and then water it. Take the root tip at 9:00-10:00 the next morning after watering, and use N 2 After O treatment for 2 hours, acetic acid with a mass percentage of 90% was fixed on ice for 10 min, and after fixation, it was washed with deionized water on ice for 10 min. Enzyme hydrolyze in a mixture of 2% cellulase at 37°C for 2 hours, wash the root tip twice with 70% ethanol by volume after enzymatic hydrolysis, then mash the root tip in 40 μL ethanol with a dissecting needle, and vortex to suspend Cells, centrifuge to retain the precipitate and add 30 μL of anhydrous acetic acid, mix well, absorb 5-8 μL droplet, place at room temperature for 5 minutes, and then perform microscopic examination. The microscopically well-examined slides are placed in a UV cross-linking instrument and passed through 120-125mJ / cm 2 Process for 2 minutes to fix the chromosomes to ob...