A kind of preparation method of Usnea pineapple nucleus suspension

A technology of Usnea pineapple and cell nucleus, which is applied in the field of preparation of Usnea pineapple cell nucleus suspension, can solve problems such as difficulty in obtaining Usnea pineapple cell nucleus suspension, cumbersome reagents in related methods, hindering research, etc., and achieves easy configuration, low cost, and ease of use. Harmful effect

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

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Problems solved by technology

[0003] Usnea pineapple leaves contain more secondary metabolites, and it is difficult to obtain a Usnea pineapple cell nucleus suspension suitable for flow cytometry analysis by the reported methods, or the related methods are cumbersome and contain harmful reagents, which hinder conduct of related research

Method used

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  • A kind of preparation method of Usnea pineapple nucleus suspension
  • A kind of preparation method of Usnea pineapple nucleus suspension

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Embodiment 1

[0019] 1. Reagent configuration

[0020] 1) Preparation of cell nucleus extract solution: Weigh 2.1g of citric acid monohydrate and 1g of polyvinylpyrrolidone K-30 into a clean 250ml beaker, add 0.5ml of Tween 20 in volume; add ultrapure water to make up to 100ml. Filter through a 0.22 micron filter and store at 4°C for later use.

[0021] 2) Fluorescent dye configuration: Weigh 25mg of propidium iodide and 25mg of ribonuclease into a clean 250ml beaker and add ultrapure water to make up to 50ml. Filter through a 0.22 micron filter and store at 4°C for later use.

[0022] 2. Extraction method of Usnea pineapple nucleus suspension

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

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Abstract

The invention discloses a preparation method for a tillandsia usuneoides nucleus suspension. The preparation method comprises the following steps: 1) selecting 50 mg of tillandsia usuneoides tender leaves, putting in a plastic culture dish, and adding 1 mL of an extracting solution, wherein the extracting solution is composed of the compositions with the following final concentration: citric acid monohydrate with the concentration of 21 g / L, polyvinylpyrrolidone K-30 with the concentration of 10 g / L and Tween20 with the volume concentration of 0.5%; and 2) chopping tillandsia usuneoides 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 tillandsia usuneoides 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 method for preparing a Usnea pineapple nucleus suspension suitable for flow cytometer analysis. Background technique [0002] Usnea pineapple (Tillandsia usuneoides L.) is a special plant that grows in the air and does not need soil. The water and nutrients needed for growth can all come from the air. Usnea pineapple has strong stress resistance and can absorb toxic and harmful substances. It has been used in environmental indication, indoor viewing, and decoration. It has broad market prospects and great development potential. However, due to the special growth mode of Usnea pineapple, there are very few related studies on molecular biology, cytology and genetic breeding, and the cultivation of new varieties is also limited. With the deepening of research, the application and basic research of plant ploidy identification, genome size determination and cell cycle using flow cytometry are of great significance to broaden the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/30G01N15/14
Inventor 金亮郁永明李小白沈福泉俞信英沈晓岚丁华侨王炜勇
Owner FLOWERS & PLANTS RES DEV CENT ZHEJIANG ACAD OF AGRI SCI
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