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

Patel, Chandrashekhar V and Gopinathan, KP (1987) Determination of trace amounts of 5-methylcytosine in DNA by reverse-phase high-performance liquid chromatography. In: Analytical Biochemistry, 164 (1). pp. 164-169.

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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, {N^6}-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 \phi X174 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: Journal Article
Additional Information: Copyright for this article belongs to the Elsevier
Keywords: HPLC techniques; 5-methylcytosine; Bombyx mori; Drosophila melanogaster; trace analysis
Department/Centre: Division of Biological Sciences > Microbiology & Cell Biology
Date Deposited: 16 May 2008
Last Modified: 19 Sep 2010 04:44
URI: http://eprints.iisc.ernet.in/id/eprint/13992

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