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A method for detecting dna damage in coelomocytes of earthworms and its application

A DNA damage and earthworm technology, which is applied in the field of cell damage detection using comet electrophoresis technology, can solve the problems of unclear imaging, easy degumming, and many steps in gel production.

Inactive Publication Date: 2019-09-27
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In order to solve the deficiencies in the prior art, the present invention provides a comet test film and a comet test method according to the present invention. This method solves the above-mentioned problems such as many steps of making glue, easy degumming, and unclear imaging. Simplify the experimental steps and improve the shooting effect of the fluorescence microscope

Method used

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  • A method for detecting dna damage in coelomocytes of earthworms and its application
  • A method for detecting dna damage in coelomocytes of earthworms and its application
  • A method for detecting dna damage in coelomocytes of earthworms and its application

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Experimental program
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Embodiment Construction

[0072] Embodiment one: the making of the slide film that is used for comet experiment of the present invention

[0073] 1. The production material of the carrier film

[0074] 1. Ordinary glass slides (Sailboat brand glass slides, products of Jiangsu Feizhou Glass & Plastic Co., Ltd., specifications and models: length and width are 25.4mm×75.6mm, thickness is 1mm-1.2mm).

[0075] 2. Home improvement glass glue (Dow Corning brand, product of Shenzhen Zhenkun Chemical Co., Ltd., product model DC-7091)

[0076] 3. Glass knife (branded by Yangjiang Shibazi, product of Yangjiang Shibazi Group Co., Ltd.).

[0077] Second, the production method of the carrier film

[0078] 1. Along the short side of the glass slide, use a glass knife to cut into 3-4mm long thin strips,

[0079] 2. Use glass glue to stick the glass strips along the long axis of the glass slide. It is required to stick 4 strips for each slide, evenly spaced from each other and distributed in the middle of the slide....

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Abstract

The invention provides a comet assay method for detecting DNA damage of earthworm coelomocytes. The method comprises production of simple glue carrying films and preparation of earthworm coelomocytes,can perform comet assay by only laying a layer of glue, and is simple, high in accuracy and high in efficiency.

Description

technical field [0001] The invention belongs to the field of cell detection, in particular to a method for detecting cell damage by using comet electrophoresis technology. Background technique [0002] Single cell gel electrophoresis (single cell gel electrophoresis Assay, SCGE), also known as Comet Assay, was invented by Ostlingn in 1984 and is a technique for detecting DNA damage in eukaryotic cells. Currently, it is used in genotoxicity tests and environmental pollution tests It has a wide range of applications related to the basic research fields of molecular epidemiology, ecopathology and DNA damage repair. The principle is: lyse the cells embedded in agarose, and then unwind and electrophoresis the DNA. If the cellular DNA breaks into pieces, comet-like structures can be seen under the microscope after staining. The main experimental steps include: preparation of single cell suspension, gel preparation and spreading, lysis, unwinding, electrophoresis, neutralization,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/447
Inventor 李岗陈列忠王彥华蔡磊明许振岚张昌鹏
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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