Method for rapid detection of in vitro DNA break damage strength by plasma

Abstract

The invention relates to a method for rapid detection of in vitro DNA break damage strength by plasma, and the method is used for determining the damage strength of plasma mutagenesis methods including atmospheric and room temperature plasma (ARTP) on DNA. The method includes the steps of: 1) treating DNA coated microspheres by plasma mutagenesis method, and setting a control group without plasma treatment; 2) performing washing, and then dyeing the DNA coated microspheres with a dye; 3) determining the fluorescence value of the dyed DNA coated microspheres by a fluorescence detector; and 4) calculating the DNA relative content of the control group and the plasma mutagenesis treated microspheres according to the fluorescence value, and judging the gene damage strength of plasma. The invention utilizes DNA coated microspheres to establish a rapid detection method for in vitro DNA damage strength so as to realize direct and effective evaluation of plasma caused damage strength to DNA.

Description

technical field [0001] The invention relates to a rapid detection method for DNA damage intensity in vitro caused by plasma, in particular to establishing a rapid detection method for DNA damage intensity in vitro by using DNA-coated microspheres. Background technique [0002] High-quality microbial strains are always the core of the biological industry. How to use efficient biological mutagenesis technology to achieve rapid optimization of strains is an important task for the biological industry. [0003] Increase the mutation rate by physical (such as X-ray, γ-ray, UV, ion beam implantation, plasma), chemical (such as alkylating agent, azide), biological (such as expression of mutS, ndk gene) and other means, which can be used in Create more genotypes in a short time. Different mutagenesis methods cause DNA damage in different ways. For example, ultraviolet (UV) mainly causes cyclobutane pyrimidine dimer (CPD) and 6-4 photoproducts (6-4PPs), thereby causing mutations from...

AI Technical Summary

Problems solved by technology

For the operating parameters of the ARTP mutagenesis system, power is usually used as a parameter, and power (P in )20-120W, gas flow (Q He )5-20slpm, irradiation distance (D) is 2-5mm, but the dose of ARTP has not been accurately measured under different operating conditions

[0007] The R&D team has qualitatively verified the damage of ARTP to DNA by means of gel electrophoresis, but so far, there is a lack of rapid detection methods for the intensity of DNA damage, which can then be used to characterize the dose of ARTP

Comet method (single-cell gel electrophoresis) is a semi-quantitative method for detecting DNA breakage damage (Kumari S, Rastogi R P, Singh K L, et al. DNA damage: Detection strategies. Excli Journal, 2008, 7:44-62. ), but its operation is complicated and can only be used for large DNA fragments

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