Abstract
HPLC analysis is important for determination of total level on DNA methylation in plants. It can be used to help characterise epigenetic changes during growth, development and stress. HPLC methods have been optimised for mammalian and microbial DNA, but not for plants. This article examines several important factors in the HPLC analysis of plant DNA methylation including extraction and purification of DNA and HPLC conditions choice by using leaves of rice seedling. The experimental results showed that RNA of nucleic acid was removed by using RNase A. This study also identified critical components of HPLC analysis. With the optimized method of HPLC conditions, the better result was achieved in the chromatogram of cytosine and 5-methylcytosine in genomic DNA acid hydrolysis. The study would offer a comprehensive guide for the stringent analysis of DNA methylation in plants.
Highlights
DNA methylation is important regulation mechanism of gene regulation and expression in biological process
high performance liquid chromatography (HPLC) analysis is important for determination of total level on DNA methylation in plants
With the optimized method of HPLC conditions, the better result was achieved in the chromatogram of cytosine and 5-methylcytosine in genomic DNA acid hydrolysis
Summary
DNA methylation is important regulation mechanism of gene regulation and expression in biological process. DNA methylation plays a key role in plant such as morphogenesis, development, stress, acclimation and adaptation [1,2]. DNA methylation is concerned with controlling the morphogenetic responses in plant tissue cultures and maintaining their genetic stability, which is of particular interest to biotechnologists. HPLC is an appropriate technique for assessing the larger-scale epigenetic effects of stress and acclimation responses in plants, while this approach does not differentiate between coding and non-coding sequences of DNA, nor detect subtle differences in the methylation status of individual genes [8]. HPLC is considered the most reliable and sensitive technique to determine total DNA methylation [9,10]. This study identifies critical components of HPLC analysis and offers a reference for the stringent analysis of DNA methylation in plants
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
