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Analysis of 5-methylcytosine in DNA: II. Gas chromatography

✍ Scribed by Aharon Razin; John Sedat

Publisher
Elsevier Science
Year
1977
Tongue
English
Weight
454 KB
Volume
77
Category
Article
ISSN
0003-2697

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✦ Synopsis

A method for analyzing 5methylcytosine in DNA by gas chromatography is described. The method is based on degradation of the DNA to its free bases by treatment with trifluoroacetic acid and gas chromatography of the trimethylsilyl derivatives of the free bases. Chromatography of microgram amounts of derivatized material is conducted at isothermal conditions using a 3% SE-30 or 2% OV-225 column. The peak areas corresponding to cytosine and S-methylcytosine are used to calculate the S-methylcytosineicytosine molar ratio in DNA. The lower limit for detection of 5methylcytosine in DNA by this method is a 5-methylcytosine/cytosine molar ratio of 0.001.

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