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Nucleic Acids Research, 1985, Vol. 13, No. 21 7639-7645
© 1985

Articles

Reduced cleavage by sodium hydroxide of methyladenine in DNA sequencing

Noriyuki Okawa, Yoshitaka Suyama* and Akira Kaji

Department of Microbiology, University of Pennsylvania School of Medicine Philadelphia, PA 19104, USA *Department of Biology, University of Pennsylvania Philadelphia, PA 19104, USA

Received August 12, 1985. Accepted October 4, 1985.

When the Maxam and Gilbert DNA sequencing method which is modified by Bencini et al. (Biotechniques Jan/Feb pp4–5, 1984) is applied to DNA containing methylated adenine in a GATC sequence, the cleavage reaction by sodium hydroxide is found to be greatly reduced in comparison to that of non-methylated adenine. Thus, a faint band in A>C reaction suggests a methyl adenine and can be used for its detection. That the faint band corresponds to a methyladenine was confirmed by Sanger sequencing of the same fragment and further by Maxam and Gilbert sequencing of the complementary strand of DNA, which was replicated in an E. coli strain either having or lacking methylation enzymes.


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