African Journal of
Agricultural Research

  • Abbreviation: Afr. J. Agric. Res.
  • Language: English
  • ISSN: 1991-637X
  • DOI: 10.5897/AJAR
  • Start Year: 2006
  • Published Articles: 6837

Full Length Research Paper

Quantitative determination of DNA Methylation in tobacco leaves by HPLC

  Feng Jia1, YunPeng Fu1, WeiQun Liu2*, Zheng Du1 1College of Tobacco Science, Henan Agricultural University, Zhengzhou 450002, People’s Republic of China. 2College of Life Sciences, Henan Agricultural University, Zhengzhou, 450002, People’s Republic of China.and YiDan Zhao2        
  1College of Tobacco Science, Henan Agricultural University, Zhengzhou 450002, People’s Republic of China. 2College of Life Sciences, Henan Agricultural University, Zhengzhou, 450002, People’s Republic of China.
Email: [email protected]

  •  Accepted: 20 October 2010
  •  Published: 31 March 2011

Abstract

 

A relatively straightforward high performance liquid chromatography (HPLC) method for determination of DNA methylation in flue-cured tobacco leaves was developed and validated. DNA methylation was measured as follows: DNA was hydrolyzed to nucleotide units in perchloric acid. The bases of cytosine and 5-methylcytosine were separated on a reversed-phase C18 column at 30°. The mobile phase used methanol: 5 mmol/L pentane sulfonate (10:90, v/v) and a flow rate of 0.7 mL/min. The absorbance of bases was measured at 273 nm. The results showed that the method was accurate, quick, and cost effective to determine DNA methylation of flue-cured tobacco.  

 

Key words: Tobacco, DNA methylation, 5- Methylcytosine, high performance liquid chromatography (HPLC).

Copyright © 2024 Author(s) retain the copyright of this article.

This article is published under the terms of the Creative Commons Attribution License 4.0

Back to Vol. 6 No. 6
Back to articles
  • Articles on Google by:
SUBSCRIBE TO RSS
SUBMIT MANUSCRIPT
CONFERENCES