Method for measuring DNA methylation content of floating fern by using high performance liquid chromatography

Abstract

The invention discloses a method for measuring DNA methylation content of floating fern by using high performance liquid chromatography. The method comprises the following steps: collecting a floating fern sample to be measured and extracting DNA in the sample to obtain a DNA sample to be measured; storing the DNA sample at the temperature of -20 DEG C for later use after the DNA sample is dissolved in buffer liquid; hydrolyzing the DNA sample to be measured and centrifuging the hydrolyzed DNA sample; taking supernatant liquid so as to obtain the solution of the sample to be measured; preparing the mixed solution of standard samples of a cytosine standard sample and a 5-methylcytosine standard sample; and measuring the DNA methylation content of the DNA sample to be measured according to peak retention time and peak areas of the mixed solution of the standard samples and the solution of the sample to be measured under the same high performance liquid chromatography measurement condition. In the method, the DNA methylation content of the floating fern is measured by using the high performance liquid chromatography; and therefore, the method has the characteristics of simplicity, low cost, fast and accurate measurement, and good quantitative analysis effect. Simultaneously the high performance liquid chromatographic technology per se is used very widely, the technology is mature relatively, and the DNA extraction from the floating fern can be implemented by using a conventional method, so the method can be popularized and applied.

Description

Method for Determination of DNA Methylation Content of Water Fern by High Performance Liquid Chromatography technical field The invention relates to a method for measuring DNA methylation content, in particular to a method for measuring DNA methylation content of water fern by high performance liquid chromatography. Background technique DNA methylation has become one of the research hotspots in molecular biology. Studies have shown that adversity (such as cold, heat stress, light, drought, salt stress, heavy metal stress) will affect the methylation level of DNA. The level of DNA methylation may be different in different tissues and different growth periods and different environments. Moreover, there is a negative correlation between gene expression activity and the degree of DNA cytosine methylation, the lower the degree of DNA methylation, the higher the gene expression activity; conversely, the higher the degree of DNA methylation, the higher the gene expression activi...

AI Technical Summary

Problems solved by technology

At present, methods for determining DNA methylation content include microarray method (DNA Microarray), gene chip, and methylation-sensitive amplified polymorphism (Mthylation-Sensitive Amplified Polymorphism, MSAP) method, methylation-sensitive spot analysis (MS- DBA) and other determination methods, but due to complex technology and high cost, it is difficult to popularize and apply in actual production

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