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J Virol. 1973 August; 12(2): 218-225
Copyright © 1973 American Society for Microbiology. All Rights Reserved.

Hydroxyapatite Chromatography and Formamide Denaturation of Adenovirus DNA

Department of Microbiology, The Wallenberg Laboratory, Uppsala University, Uppsala, Sweden

ABSTRACT

Denatured adenovirus DNA was retained by hydroxyapatite columns under conditions generally used for selective retention of double-stranded DNA, probably due to several partially complementary sequences within single-stranded DNA. It was found that addition of formamide reduced the fraction of sonically treated, denatured adenovirus DNA bound to hydroxyapatite from about 30% to less than 1%. This led to a study of the effect of formamide on the melting temperature (T_m) of double-stranded DNA in solution or bound to hydroxyapatite. The T_m of DNA decreases 0.56 C/1% formamide, a value determined in buffered solutions with purified formamide.