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Soybean cell nucleus dissociation liquid suitable for flow cytometry analysis and application thereof

A flow cytometer and cell nucleus technology, applied in the biological field, can solve the problems of non-appearance, low efficiency of separating cell nuclei, unsuitable for large-scale sample preparation and detection, etc., to achieve reliable experimental results, good dissociation effect, and good dissociation effect of effect

Pending Publication Date: 2020-08-07
YANGZHOU UNIV
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Problems solved by technology

[0005] However, no nuclei dissociation solution suitable for leaf and root tissues of different soybean varieties has appeared in the prior art, and the methods of enzymatic hydrolysis and mechanical cutting used in other species to separate nuclei have low efficiency and are not suitable for large-scale samples Preparation and detection

Method used

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  • Soybean cell nucleus dissociation liquid suitable for flow cytometry analysis and application thereof
  • Soybean cell nucleus dissociation liquid suitable for flow cytometry analysis and application thereof
  • Soybean cell nucleus dissociation liquid suitable for flow cytometry analysis and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] 1 Materials and methods

[0027] 1.1 Experimental materials

[0028] The tested materials are soybean varieties including Williams 82, Holladay, PI 567611, PI 567651, and Fiskeby III, which are preserved by the soybean genetics and breeding laboratory of Yangzhou University.

[0029] 1.2 Method

[0030] 1.2.1 Plant cell nuclei dissociation solution preparation

[0031] The formulations of the six nuclear dissociation solutions used in this experiment are listed in Table 1.

[0032] Table 1 Nuclei Dissociation Buffer Recipe

[0033]

[0034]

[0035] Avoid repeated freezing and thawing of the nuclear dissociation solution, and it can be stored at 4°C for several days after thawing.

[0036] 1.2.2 PEG 6000 simulated drought treatment

[0037] Soybean seeds were planted in the soil after absorbing water and swollen, and germinated and grown for 6 days. Bean sprouts with the same growth state were selected, and the dust was washed under the premise of avoiding da...

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Abstract

The invention belongs to the field of biology, and particularly relates to a soybean cell nucleus dissociation solution suitable for flow cytometry analysis and application thereof. The soybean cellnucleus dissociation liquid suitable for flow cytometry analysis comprises 45mM magnesium chloride, 20mM MOPS, 30 mM of sodium citrate and 30 mM of a non-ionic surface active agent; the nuclear dissociation liquid provided by the invention has good applicability on different soybean varieties, not only has a good nuclear dissociation effect on soybean leaves and root tissues, but also is reliablein corresponding cell cycle analysis experiment result, and provides technical support for further researching the influence of adversity on the soybean intrakaryotic replication level; and a foundation is laid for simply, conveniently, quickly and accurately researching the content, the cell cycle and the genetic ploidy of the soybean cell nucleus DNA by using a flow cytometry.

Description

technical field [0001] The invention belongs to the field of biology, and in particular relates to a soybean nucleus dissociation solution suitable for flow cytometer analysis and application thereof. Background technique [0002] Flow cytometry is a method for multi-parameter, rapid analysis, quantification and sorting of single cells or biological particles in liquid flow using flow cytometry. With the development of fluorescent molecular labeling technology, flow cytometry is widely used in the field of life sciences: such as analyzing various metabolic characteristics of cells by flow cytometry, judging cell apoptosis and cell necrosis, and evaluating cell differentiation and proliferation properties, as well as DNA content analysis, determination of cell population proportions, and sorting. The application of flow cytometry in botany research mainly includes the following aspects: (1) Plant cell nucleus analysis, such as: determination of DNA content, plant genetic plo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G01N1/30
CPCG01N1/28G01N1/30
Inventor 宋丽陈林戴毅潘贞志
Owner YANGZHOU UNIV
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