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

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.

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...

AI Technical Summary

Problems solved by technology

However, during the operation of FISH technology, the quality of chromosome slide specimens, the method and efficiency of probe labeling, the temperature and time of co-denaturation of probes and chromosomes all affect the experimental results. For example, the quality of chromosome slide specimens determines whether FISH can hybridize. One of the key factors for success, insufficient or excessive enzymatic hydrolysis will lead to the failure of chromosome preparation; the cytoplasm of the slide is too dense, the probe is not easy to hybridize and will cause high background; whether the pretreatment is appropriate determines whether the chromosome length is moderate, whether there is conducive to hybridization

Different methods of probe labeling will also affect the hybridization results accordingly

The temperature and time of the co-denaturation of the probe and the chromosome are also important factors. If the co-denaturation temperature is low or the time is short, the double strands are not completely opened, and no specific signal will be hybridized. However, if the co-denaturation temperature is too high or the time is too long, the chromosome Expansion and deformation, making the experimental results abnormal

FISH technology has been successfully applied to many plants, but there is no report on the application of fluorescence in situ hybridization technology in Asparagus asparagus

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