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Nucleic Acids Research, 1978, Vol. 5, No. 5 1479-1485
© 1978

Articles

Detection of 5-methylcytosine in DNA sequences

Haruo Ohmori, Jun-ichi Tomizawa and Allan M. Maxam*

Laboratory of Molecular Biology, National Institute of Arthritis, Metabolism and Digestive Diseases, National Institutes of Health Bethesda, MD 20014, USA

Received March 13, 1978.

Col E1 DNA has methylated cytosine in the sequence 5'-CC*(A/T)GG-3' and methylated adenine in the sequence 5'-GA*TC-3' at the positions indicated by asterisks(*). When the Maxam-Gilbert DNA sequencing method is applied to this DNA, the methylated cytosine (5-methylcytosine) is found to be less reactive to hydrazine than are cytosine and thymine, so that a band corresponding to that base does not appear in the pyrimidine cleavage patterns. The existence of the methylated cytosine can be confirmed by analyzing the complementary strand or unmethylated DNA. In contrast, the methylated adenine (probably N6-methyladenine) cannot be distinguished from adenine with standard conditions for cleavage at adenine.

*Department of Biochemistry and molecular Biology, Harvard University, Cambridge, MA 02138, USA


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