Fluorescence in situ hybridization method for high resolution chromosomes of cucumber

Abstract

The invention provides a fluorescence in situ hybridization method, wherein fluorescence in situ hybridization with multiple probes is performed based on chromosomes in the meiosis pachytene stage of cucumber as a target DNA (Deoxyribonucleic Acid). The method comprises the following steps of: selecting high quality films with good chromosome dispersion and clear morphological structure, degenerating in 70% deionized formamide at 80 DEG C, and dehydrating and drying in gradient alcohol; extracting a target plasmid DNA by alkaline lysis, marking by an incision translation method, preparing a hybrid mixed liquor, degenerating at 90 DEG C; and loading to a glass slide to hybridize the target DNA and probe; and finally detecting signals. The fluorescence in situ hybridization technology provided by the invention can be applied to drawing a physical map of chromosomes of cucumber. As multiple probes can be added at one time, repeated hybridization to one film can be avoided, so that the experimental efficiency is greatly improved. According to the invention, appropriate whole genome DNA and Cot-1DNA of cucumber are added in the process of configuring the hybrid mixed liquor for blocking spurious signals and repetitive sequence signals, so that the identification rate of target signals is improved and the accuracy of experimental result is increased.

Description

1. Technical field [0001] The invention discloses a fluorescence in situ hybridization method of cucumber high-resolution chromosomes, which is specially used for the fluorescence in situ hybridization of cucumber pachytene chromosomes, is beneficial to the development of cucumber cytology research, and belongs to the field of plant biotechnology. 2. Technical background [0002] Cucumber (Cucumis sativus L., 2n=14), also known as cucumber and cucumber, is a vine herbaceous plant of Cucurbitaceae Melon. Because of its delicious fruit, rich nutrition, and the effects of anti-tumor, anti-aging, weight loss, strengthening the body, strengthening the brain and calming the nerves, it is deeply loved by consumers. [0003] Invention and use of cucumber pachytene chromosomes as target DNA for fluorescence in situ hybridization is an important progress in the field of cytology in recent years. At present, there are only a few reports of fluorescent in situ hybridization using the p...

AI Technical Summary

Problems solved by technology

For the previous fluorescent in situ hybridization using the interphase nucleus or metaphase chromosome of cucumber as the target DNA, due to the small size of the cucumber chromosome, the high chromatin concentration of the metaphase chromosome, and the thick and short chromosomes, the recognition rate of the signal positioned on it is not high; On the other hand, due to the interference of background signals or repetitive sequence signals during the hybridization process, it will adversely affect the detection of target signals

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