The effects of some restriction endonucleases (REs) on the nucleolar organizing regions and on the genes for ribosomal RNA (rDNA) were analyzed using the nucleolar organizing region of the chromosome 6 of Tapinoma nigerrimum as an experimental model, since, in accordance with previous studies, the genes for ribosomal RNA seem to be present only in this chromosome. In situ non-digestion of the nucleolar organizing region was observed when EcoRI and HindIII were used. However, very evident digestion and partial digestion respectively were observed when HaeIII and Tru9I were used. Southern blot analysis realized on naked DNA digested with the same REs and using rDNA of Drosophila melanogaster as probe showed that there are target sequences for these enzymes in the rDNA. In accordance with the results obtained, the rDNA is poor in EcoRI and HindIII sequences, contains moderate amounts of Tru9I sequences and is rich in HaeIII sequences. All the data obtained suggest that in the nucleolar organizing region of Tapinoma nigerrimum, the major, if not the only, limiting factor affecting in situ digestion by the REs used is the presence and frequency of their specific restriction targets. Consequently, extraction of DNA from this chromosome region depends on the size of the fragments originated.