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Fluorescence in situ hybridization method for metaphase chromosomes of Zanthoxylum bungeanum

A technique of fluorescence in situ hybridization and Zanthoxylum bungeanum, which is applied in the fields of cytogenetics and molecular biology, can solve the problems of difficult seed germination, difficult clipping of root tips, long time, etc., and achieves less probe DNA and less probe Ease of preparation and improved detection efficiency

Inactive Publication Date: 2020-08-21
SICHUAN AGRI UNIV
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Problems solved by technology

The main reasons are: first, the seeds of Zanthoxylum bungeanum are not easy to germinate, and the time is long.
Second, the root tip tissue is rich in oil, which seriously affects the observation of chromosomes under the microscope and is not conducive to finding chromosomes
Third, Zanthoxylum plants have many and small chromosomes and are not easy to disperse, which is not conducive to finding a cleavage phase with a complete number of chromosomes in the metaphase
FISH technology has been successfully applied to many plants, but there is no application report on any species of Zanthoxylum, including Zanthoxylum bungeanum

Method used

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  • Fluorescence in situ hybridization method for metaphase chromosomes of Zanthoxylum bungeanum

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Embodiment Construction

[0020] The probe sets used in the implementation of the present invention are all synthesized by Beijing Sangong Company. The synthesized probes are diluted with 1×TE to a concentration of 100M and stored at -20°C. Store at ℃.

[0021] The present invention will be described in further detail below in conjunction with the accompanying drawings and specific embodiments, which are explanations rather than limitations of the present invention.

[0022] A specific embodiment of the present invention provides a method for fluorescent in situ hybridization of metaphase chromosomes of Zanthoxylum bungeanum, comprising the following steps:

[0023] 1) Preparation of mid-stage chromosome slide specimens of Zanthoxylum bungeanum: Transplant the annual Zanthoxylum bambooi plant grown in the field into a pot filled with nutrient soil, and cultivate it at room temperature for two months until the root tip grows to 1.5 ~2.0cm, cut 1~1.5cm root tip tissue at 10:30 am and 4:30 pm, and use N ...

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Abstract

The invention provides a fluorescence in situ hybridization method for zanthoxylum armatum metaphase chromosome. The fluorescence in situ hybridization method comprises the steps of zanthoxylum armatum metaphase chromosome glass slide specimen preparation, fluorescence in situ hybridization, signal detection and glass slide recovery. The fluorescence in situ hybridization method disclosed by the invention is beneficial to establishing a stable and convenient metaphase chromosome fluorescence in situ hybridization reaction system, greatly improves the detection efficiency of the zanthoxylum armatum metaphase chromosome and provides a novel method for detecting the anthoxylum armatum chromosome and even zanthoxylum species chromosome.

Description

technical field [0001] The invention belongs to the field of cytogenetics and molecular biology, and in particular relates to a fluorescence in situ hybridization method for metaphase chromosomes of Zanthoxylum bungeanum. Background technique [0002] Existing methods for detecting plant chromosomes generally use plant chromosome banding, which can be divided into two categories, namely fluorescence banding and Giemsa banding. But neither method has been used in Zanthoxylum genus. The main reasons are: first, Zanthoxylum bungeanum seeds are not easy to germinate, and the time is long, and the seed shell is hard, so it is not easy to absorb water and swell, and it is difficult to use the seeds to germinate and cut the root tip. Second, the root tip tissue is rich in oil, which seriously affects the observation of chromosomes under the microscope and is not conducive to finding chromosomes. Third, Zanthoxylum plants have many and small chromosomes and are not easy to dispers...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895
Inventor 罗小梅陈亮万文林龚伟王景燕刘俊成
Owner SICHUAN AGRI UNIV
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