A kind of preparation method of strawberry cell nucleus suspension

A cell nucleus and strawberry technology, which is applied in the field of preparation of strawberry nucleus suspension, can solve problems such as obstacles to research, difficulty in obtaining strawberry nucleus suspension, cumbersome related methods and harmful reagents to the human body, and achieve low cost, easy configuration, and time-saving effects

Active Publication Date: 2016-01-20
ZHEJIANG XIAOSHAN COTTON & FLAX RES INST
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Strawberry leaves contain polyphenols and other secondary metabolites. It is difficult to obtain a strawberry 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 conduct of research

Method used

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

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Embodiment 1

[0019] 1. Reagent configuration

[0020] 1) Cell nucleus extract preparation: Weigh 1.47g trisodium citrate dihydrate and 1g polyvinylpyrrolidone K-30 into a clean 250ml beaker, add 0.5ml Tween 20 at a volume concentration; 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. Strawberry cell nucleus suspension extraction method

[0023] (1) Take young strawberry leaves (about 50mg), put them in a plastic petri dish and add 1ml 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 a suspension of strawberry nuclei; (4) Add DNA fluorescent dye iodine to the filtered extract Propid...

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Abstract

The invention discloses a method for preparing strawberry cell nucleus suspension. The steps are as follows: 1) Take 50 mg of young strawberry leaves, put them in a plastic petri dish and add 1 ml of the extract, which is mainly composed of the following components at the final concentration: 14.8 g / L trisodium citrate dihydrate, 10 g / L Polyvinylpyrrolidone K-30, and Tween 20 at a volume concentration of 0.5%; 2) Use a blade to chop young strawberry leaves in the extract, then collect the extract and filter it through a 350-mesh nylon mesh to obtain a suspension of strawberry nuclei. The beneficial effects of the present invention: 1) The use of harmful reagents, such as mercaptoethanol, is reduced to the greatest extent, and the harm of the extract to the human body is reduced; 2) The extract has simple ingredients, only three kinds, all of which are conventional laboratory reagents, and the cost Inexpensive, easy to configure, can greatly reduce the cost; 3) The steps are simple, no centrifugation is required, which saves time and improves efficiency, and can obtain high-quality cell nucleus suspension for flow cytometry detection in a short time.

Description

technical field [0001] The invention relates to a method for preparing strawberry cell nucleus suspension suitable for flow cytometer analysis. Background technique [0002] Cultivated strawberries ( Fragaria×ananassa Duch.), also known as pineapple strawberry, belongs to the Rosaceae strawberry genus perennial herb, the fruit is bright in color, soft and juicy, rich in nutrients such as sugar, protein, vitamins and anthocyanins, and is an important economic crop widely cultivated in the world , one of the world's seven major fruits. It is generally believed that the genetic background of current strawberry cultivars is narrow, and the development of new varieties is thus 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 basis of strawberry breeding. Among them, obtaining strawberry cell nucleus suspension suitabl...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/28
Inventor 金亮李小白王炜勇马广莹刘慧春邹清成
Owner ZHEJIANG XIAOSHAN COTTON & FLAX RES INST
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