Abstract
The chromatographic behavior of DNA denatured by heat at various temperatures, and reacted or not with formaldehyde, on hydroxyapatite columns has been investigated using both stepwise and gradient elution. The recovery of denatured DNA from the columns and the behavior of the chromatographic fractions upon rechromatography have been studied. The bulk of denatured DNA from bacteria and higher animals is eluted from hydroxyapatite columns by a phosphate molarity distinctly lower than that eluting native DNA, whereas a small “native-like” fraction is eluted at the same molarity as native, renatured, and denatured, cross-linked DNA. Investigations on the ultraviolet absorbance-temperature profile and formaldehyde reaction of the “native-like” fraction showed that it is endowed, to a large extent, with complementary base pairing. Subsequent work has shown that the “native-like” fraction is the carrier of the residual transforming activity surviving denaturation, in the case of bacterial transforming DNA; whereas, it is formed by satellite DNA in the case of DNA's from higher animals. Denatured, single-stranded DNA molecules may be fractionated on hydroxyapatite columns according to their base composition.
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