Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Cell DNA damage detection method

A technology for DNA damage and detection methods, which is applied in the direction of measuring devices, instruments, and material analysis through electromagnetic means, can solve the problems of sophisticated and complex production processes, high hardware and technical requirements, and achieve simplified preparation processes and improved efficiency , the effect of not easy cell overlap

Active Publication Date: 2014-04-30
XIAMEN UNIV
View PDF0 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The microchamber array method improves the preparation process of the glue plate compared with the conventional SCGE, and the position of each cell in the microchamber is clear, and automatic analysis is used to ensure that every cell is analyzed. However, the microchamber mold manufacturing process in the microchamber array method is precise. Complicated, the mold needs to change with the size of the cells. When changing the tested cells, the mold needs to be re-optimized. The process is complicated, and automatic image reading requires special supporting equipment. The hardware and technical requirements are high, which affects its promotion and application.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Cell DNA damage detection method
  • Cell DNA damage detection method
  • Cell DNA damage detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] (1) Preparation of splenocyte samples from mice (Cavia porcellus): kill the mice by cervical dislocation, and quickly remove the spleen in an ultra-clean bench after being briefly treated with 75% ethanol, place it in a glass plate, and wash it off with pre-cooled PBS buffer After removing the blood on the surface, immerse it in PBS buffer, and cut it into 1-2mm pieces with ophthalmic surgical scissors 3 of small pieces. Inhale the PBS buffer with the spleen fragments suspended into a disposable sterile syringe (with the needle removed) and press-filter repeatedly, pass through a 300-mesh cell sieve, and collect the cell suspension. At room temperature, centrifuge at 1500rpm for 10min to get the cell pellet, then resuspend it in RMPI-1640, and adjust the cell density to about 8×10 5 individual / mL.

[0036] (2) Cell contamination: seed the cells in a 24-well cell culture plate, and then add H 2 o 2 solution, H 2 o 2 The concentrations were 1 μmol / L, 20 μmol / L, and ...

Embodiment 2

[0046] (1) Preparation of human acute mononuclear cell (THP-1) samples: THP-1 cells were cultured in RMPI1640 medium with 10% fetal bovine serum, 100 U / mL penicillin, and 100 U / mL streptomycin at 37°C for 5 %CO2 cell incubator culture. When passaging, pipette gently with a pipette gun to make the cells uniform and then pass directly. According to the needs of the experiment, the cells are seeded in a 24-well cell culture plate and cultured for 24 hours before use.

[0047] (2) Cell contamination: seed the cells in a 24-well cell culture plate, then add HgCl 2 solution, HgCl 2 Concentrations were 1 μmol / L, 10 μmol / L, 100 μmol / L, and a blank control group was set up at 4°C for 1.5 hours. After the pollution, wash twice with PBS, collect the cells by centrifugation at 1200rpm, resuspend with PBS, and adjust the cell density to about 8×10 5 pc / mL for use.

[0048] (3) Subsequent operations refer to steps 3-9 of Example 1. See the experimental results figure 2 .

Embodiment 3

[0050] (1) For the preparation of mouse (Cavia porcellus) splenocyte samples, refer to Step 1 of Example 1.

[0051] (2) Cell contamination: seed the cells in a 24-well cell culture plate, then add CuCl 2 solution, CuCl 2 Concentrations were 10nmol / L, 100nmol / L, 1μmol / L respectively, and a blank control group was set up at 4°C for 1.5h. After the pollution, wash twice with PBS, collect the cells by centrifugation at 1000rpm, resuspend with PBS, and adjust the cell density to about 8×10 5 pc / mL for use.

[0052] (3) Subsequent operations refer to steps 3-9 of Example 1. See the experimental results image 3 .

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a cell DNA damage detection method, and relates to the cell damage detection. The cell DNA damage detection method comprises the following steps: preheating a to-be-detected cell sample with low-gelling temperature agarose prepared from calcium and magnesium-free PBS (Phosphate Buffer Solution) to obtain a mixed liquid, paving the mixed liquid in a plate cover sample hole of a 24-hole cell culturing plate, spreading agarose evenly, cooling, curing, and then immersing the plate cover into cell lysis buffer, and pyrolyzing; washing an agarose plate after pyrolysis by an alkaline unwinding liquid, and then immersing the agarose plate into a container filled with a precooling alkaline unwinding liquid; placing the agarose plate after alkaline unwinding into an electrophoresis tank filled with a precooling electrophoresis liquid for electrophoresis; immersing the agarose plate after the electrophoresis into milli-Q water; standing and immersing the agarose plate by precooling absolute ethyl alcohol, and refrigerating and storing the taken agarose plate; adding a drop of MillI-Q water dropwise in each hole of the agarose plate after dehydration and drying, standing, wetting the agarose plate, adding 10-15micron L of ethidium bromide dropwise in each hole, keeping away from light, dyeing, sucking residual dye liquid dry on the surface of the agarose plate by filtration paper, and collecting images with excitation wavelength being 515nm.

Description

technical field [0001] The invention relates to cell damage detection, in particular to a cell DNA damage detection method. Background technique [0002] In cancer patients, the stability of DNA genetic information is damaged to varying degrees, which indicates that the occurrence of cancer is closely related to DNA damage. There are various harmful physical and chemical factors in the environment (such as ultraviolet radiation, pesticide residues and other organic pollutants, etc.) that can directly or indirectly cause cell DNA damage, and a large number of new factors enter the environment every year, so Rapid and effective detection of DNA damage and toxicity of existing and newly entered environmental factors is conducive to taking timely measures to control and treat harmful factors, reducing the occurrence of related cancers and genetic diseases, and is of great significance for environmental monitoring and protection. [0003] Single cell gel electrophoresis (Single ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N21/63G01N27/447
Inventor 韩大雄肖丹王海燕
Owner XIAMEN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com