Cross-linking of double-helical nucleic acids with a photoreactive analogue of ethidium

Intercalation of the ethidium analogue 3,5-diazido-5-ethyl-6-phenylphenanthridinium into double helices followed by irradiation with blue or ultraviolet light results in cross-linking between the two strands with an efficiency around 30% for DNA, RNA, and DNA-RNA hybrids. Details of this reaction and a convenient synthesis of the ethidium analogue are described. Stable tertiary structure in RNA impedes intercalation and thus reduces the efficiency of cross-linking. In contrast to the ethidium derivative, various acridine diazides show little or no cross-linking ability.