Direct incorporation of fluorescein-12-dUTP to insect fixed chromosomes by random primed extension.

The use of an in situ random primed system allows the direct incorporation of fluorescein-12-dUTP into fixed insect chromosomes, resulting in a strong fluorescent labelling. While in an orthopteran species (Eyprepocnemis plorans) a relatively uniform labelling of meiotic and mitotic chromosomes is produced, in Tenebrio molitor (Coleoptera) only the chromosomal arms, but not the pericentromeric heterochromatic areas of mitotic chromosomes, show positive labelling by this method. This indicates that the organization of DNA in heterochromatin is distinct from that in the euchromatin in distantly related species, and in such a way that, in some species, the random hexanucleotides are prevented from annealing with the chromosomal denatured DNA or (and) that primer extension by Klenow enzyme is impeded.