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A kind of preparation method of anthurium cell nucleus suspension

A technology for cell nucleus and anthurium, which is applied in the field of preparation of anthurium cell nucleus suspension, can solve the problems of complicated reagents in related methods, difficulty in obtaining anthurium cell nucleus suspension, hindering research, etc., and achieves the effects of low cost, easy configuration and harm reduction.

Active Publication Date: 2016-08-24
FLOWERS & PLANTS RES DEV CENT ZHEJIANG ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Anthurium leaves contain many secondary metabolites, and it is difficult to obtain an anthurium cell nucleus suspension suitable for flow cytometry analysis by the reported methods, or the related methods are cumbersome and contain harmful reagents, which hinder the relevant research carry out

Method used

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  • A kind of preparation method of anthurium cell nucleus suspension
  • A kind of preparation method of anthurium cell nucleus suspension

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Embodiment 1

[0019] 1. Configuration of reagents

[0020] 1) Configuration of cell nucleus extract: Weigh 2.1g of citric acid monohydrate and 1g of polyvinylpyrrolidone K-40 in a clean 250ml beaker, and add ultrapure water to make the volume to 100ml. After filtering with a 0.22 micron filter membrane, store at 4°C for later use.

[0021] 2) Fluorescent dye configuration: Weigh 25mg propidium iodide and 25mg ribonuclease into a clean 250ml beaker and add ultrapure water to make the volume to 50ml. After filtering with 0.22 micron filter membrane, store at 4°C for later use.

[0022] 2. Extraction method of Anthurium andraeanum cell nuclear suspension

[0023] (1) Take the young leaves of Anthurium andraeanum (about 50 mg), place them in a plastic petri dish and add 1 ml of extract. (2) Use a sharp razor blade to chop the leaves in the extract; (3) Collect the extract and filter it through a 350-mesh nylon mesh to obtain the nucleus suspension of Anthurium andraeanum; (4) Add DNA fluorescent dye ...

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Abstract

The invention discloses a preparation method for anthurium andraeanum cell nucleus suspension. The preparation method comprises the following steps: 1) taking 50 mg of anthurium andraeanum tender leaves, putting in a plastic culture dish, adding 1 mL of an extracting solution, wherein the extracting solution is mainly composed of the compositions with the following final concentration: citric acid monohydrate with the concentration of 21 g / L and polyvinylpyrrolidone K-40 with the concentration of 10 g / L; and 2) chopping anthurium andraeanum tender leaves in the extracting solution by employing a blade, collecting the extracting solution and filtering with a 350-mesh nylon net, so as to obtain the anthurium andraeanum cell nucleus suspension. The beneficial effects comprise that 1) usage of harmful reagents, such as mercaptoethanol, are furthest minimized, and harm of the extracting solution on human body is mitigated; 2) the extracting solution is simple in compositions, and the compositions comprise three kinds and are all routine laboratory reagents, the cost is low, the preparation is simple, and the cost is substantially reduced; and 3) the steps are simple, centrifugation is not needed, time is saved, efficiency is improved, and the high-quality cell nucleus suspension can be obtained in a short time for flow cytometer detection.

Description

Technical field [0001] The invention relates to a preparation method of an Anthurium andraeanum cell nuclear suspension suitable for flow cytometry analysis. Background technique [0002] Anthurium (Anthurium andraeanum), its flowers are unique, with buddha flame inflorescences, bright and gorgeous, rich in color, leaf-shaped bracts, common bracts are red, pink, white, etc., which have great ornamental value and are the world Famous flowers. It is generally believed that the genetic background of current cultivated varieties of Anthurium andraeanum is narrow, and the cultivation of new varieties is therefore restricted. With the deepening of research, the use of flow cytometry to identify the ploidy of anther culture plants, determine the genome size and cell cycle and other applications and basic research is of great significance to broaden the foundation of Anthurium andraeanum breeding. Among them, obtaining a nuclear suspension of Anthurium andraeanum cells suitable for flo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/28
Inventor 金亮田丹青李小白葛亚英刘建新沈福泉王炜勇
Owner FLOWERS & PLANTS RES DEV CENT ZHEJIANG ACAD OF AGRI SCI
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