TITLE:
Research of total levels on DNA methylation in plant based on HPLC analysis
AUTHORS:
Qiang Chen, Siyuan Tao, Xiaohua Bi, Xin Xu, Lanlan Wang, Xuemei Li
KEYWORDS:
DNA Methylation; Cytosine; 5-Methylcytosine; High Performance Liquid Chromatography; Mobilephase
JOURNAL NAME:
American Journal of Molecular Biology,
Vol.3 No.2,
April
30,
2013
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.