Determination of trace amounts of 5-methylcytosine in DNA by reverse-phase high-performance liquid chromatography

Patel, Chandrashekhar V. ; Gopinathan, K. P. (1987) Determination of trace amounts of 5-methylcytosine in DNA by reverse-phase high-performance liquid chromatography Analytical Biochemistry, 164 (1). pp. 164-169. ISSN 0003-2697

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Official URL: http://linkinghub.elsevier.com/retrieve/pii/000326...

Related URL: http://dx.doi.org/10.1016/0003-2697(87)90381-2

Abstract

A high-performance liquid chromatographic method to separate five major bases (cytosine, thymine, guanine, adenine, and uracil) and three minor methylated bases (5-methylcytosine, N6-methyladenine, and 7-methylguanine) has been developed using a volatile mobile phase under isocratic conditions. It is extended to quantitate 5-methylcytosine in trace amounts (1 in 20,000 cytosine residues). The suitability of the method has been verified by estimating 5-methylcytosine in DNAs of φX174 and pBR322. The method has been applied to quantitate the extent of cytosine methylation in DNA of larval silk glands of Bombyx mori. Our results confirm that the pupal DNA of Drosophila melanogaster does not contain detectable amounts of 5-methylcytosine.

Item Type:Article
Source:Copyright of this article belongs to Elsevier Science.
Keywords:HPLC Techniques; 5-methylcytosine; Bombyx mori; Drosophila melanogaster; Trace Analysis
ID Code:23628
Deposited On:26 Nov 2010 08:43
Last Modified:02 Jun 2011 06:13

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