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Simultaneous R-banding and localization of dA-dT clusters in human chromosomes

2008; BioMed Central; Volume: 98; Issue: 2 Linguagem: Inglês

10.1111/j.1601-5223.1983.tb00601.x

1601-5223

M Rønne,

Chromosomal and Genetic Variations

In vitro exposure of different cell lines to dA-dT probes is known to induce species-specific modifications in metaphase chromosomes. In human lymphocytes the main measurable effect is a general inhibition of chromosome contraction. The inhibition is pronounced in specific regions of the human karyotype, so these regions appear elongated. Simultaneous in vitro exposure of human PHA-stimulated lymphocytes to BUdR and one of the dA-dT specific agents distamycin A or Hoechst 33258 for 6.5 h prior to harvest induces R-banded chromosomes both after conventional Giemsa and FPG-staining. Because of the banding the position of the most affected regions can be precisely determined. Thus the combined in vitro exposure to BUdR and one of the dA-dT specific agents offers a simple and precise means to localize major dA-dT clusters in mammalian genomes with traditional Giemsa staining, and provides after FPG-staining high resolution R-bands.