Comet assay method and device for DNA damage of sugarcane leaves

A DNA damage and comet test technology, applied in DNA preparation, recombinant DNA technology, tissue cell/virus culture devices, etc., to achieve the effect of high detection sensitivity and improved efficiency

Pending Publication Date: 2022-04-15
SUGARCANE RES INST OF YUNNAN ACADEMY OF AGRI SCI
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Problems solved by technology

[0005] In order to make up for the deficiencies of the prior art and solve the problem that the comet test method has not been used to c

Method used

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  • Comet assay method and device for DNA damage of sugarcane leaves
  • Comet assay method and device for DNA damage of sugarcane leaves
  • Comet assay method and device for DNA damage of sugarcane leaves

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Embodiment approach

[0052] As an embodiment of the present invention, in said S1, the preparation of tissue cells includes the following steps:

[0053] A1: Use clean scissors to cut vigorously growing fresh sugarcane+1 leaf, and take about 500mg of leaf tissue with appropriate length and width;

[0054] A2: Place the culture dish obliquely on the fixed base in the fixing mechanism, add pre-cooled 1ml 1×PBS (no Ca2+Mg2+, 20mM EDTA) solution at the base of the culture dish;

[0055] A3: Invade one end of the blade into the PBS solution, and cut it quickly with a sharp scalpel blade, as thin as possible, and keep the cut surface in contact with the solution;

[0056] A4: Wash the sectioned leaf tissue with a small amount of PBS solution, let it stand for 2 minutes, use a pipette to absorb the tissue cell liquid, and filter through a 200-mesh sieve to remove tissue fragments;

[0057] A5: The cell nucleus suspension was further enriched by centrifugation at 1500g, 4°C for 10 minutes.

[0058] As a...

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Abstract

The invention belongs to the technical field of molecular biology, and particularly relates to a comet assay method and device for DNA (deoxyribonucleic acid) damage of sugarcane lamins.According to the method, a large number of complete sugarcane lamina cell nucleuses can be obtained, so that the statistical magnitude required by the comet assay can be guaranteed; in order to obtain a good comet assay result, the premise is to obtain a large number of plant cell nucleuses, and the larger the statistical number in comet assay result analysis is, the more reliable the result is generally; the number of the cell nucleuses prepared in the steps S1 to S5 can reach 106/ml, and the experimental requirements are fully met; meanwhile, the change of the DNA damage of the sugarcane leaves can be well reflected by the experimental conditions established by the method disclosed by the invention; by comparing the experimental group and the control group, the change of the comet shape can be intuitively observed, and the detection sensitivity is very high, so that the method is favorable for being popularized to the change rule of DNA damage in numerous life activities such as apoptosis of sugarcane leaves, adversity stress response and diseases.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, in particular to a comet test method and device for DNA damage of sugarcane leaves. Background technique [0002] The integrity of biological genome DNA molecules is closely related to the normal life activities of cells. Stress from external conditions and apoptosis can lead to DNA damage; DNA damage is likely to affect the accuracy of DNA replication, transcription and translation, disturb the expression and regulation of normal genes, and also produce mutagenic proteins that lead to cell mutation ; Therefore, DNA damage detection and evaluation technology is conducive to clarifying the laws of many life activities such as cell apoptosis, stress response and disease, and plays an important role in disease diagnosis, evaluation and environmental protection. [0003] The basic principle of DNA damage detection is that DNA damage will lead to changes in the physical and chemical propert...

Claims

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Application Information

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IPC IPC(8): C12Q1/6806C12Q1/6895C12N15/10C12M1/22C12M1/02C12M3/00
Inventor 胡鑫刘少春高欣欣武晋宇刘高源郭家文邓军李如丹樊仙贾迪迩
Owner SUGARCANE RES INST OF YUNNAN ACADEMY OF AGRI SCI
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