It is thought that statistical analysis of dinucleotide periodicities can provide insight into the general features of nucleosome forming sequences. The chromatin of simian virus 40 (SV40) provides a model for a unique DNA sequence that is found in association with histones in vivo. I have therefore analyzed the periodicity of dinucleotides in a collection of cloned nucleosomal DNA fragments prepared from SV40 chromatin isolated under relatively mild conditions, in order to learn about the generality of results obtained from the statistical approach and to examine the SV40 data set in the context of models that have been proposed to explain the molecular basis of nucleosome formation. In one study, I assumed a symmetry in the distribution of dinucleotides with respect to the nucleosome dyad position and considered complementary dinucleotides to be equivalent, i.e. AA = TT and GG = CC. The results showed a periodic signal for GG/CC but not for AA/TT, purine-purine, and pyrimidine-pyrimidine dinucleotides. In a second study, the SV40 nucleosomal DNA fragments were aligned and examined with respect to the late strand of the viral genome to determine the distribution of dinucleotides in one direction. Fourier analysis revealed periodic signals for AA/TT (10.26 bp) and GG/CC (10.0 bp) and indicated that AA dominates the occurrences of AA/TT and GG dominates the occurrences of GG/CC. The results of both studies implied that there might be an asymmetry and a directionality in the distribution of certain dinucleotides in nucleosomes.