Detection of species specific chromosomes in somatic cell hybrids

• Published in: Somatic cell and molecular genetics 1985-11, Vol.11 (6), p.571-577
• Main Authors: DURNAM, D. M, GELINAS, R. E, MYERSON, D
• Format: Article
• Language: English
• Subjects: Animal cells; Animals; Biological and medical sciences; Biotechnology; Bone Marrow - enzymology; Bone Marrow Cells; Cell Line; Chromosomes - ultrastructure; Cricetinae; Cytogenetics; DNA - genetics; DNA - isolation & purification; Eukaryotic cell cultures; Female; Fundamental and applied biological sciences. Psychology; Genetical aspects; Genetics of eukaryotes. Biological and molecular evolution; Humans; Hybrid Cells - cytology; Hybrid Cells - enzymology; Kidney; Methods and miscellaneous; Methods. Procedures. Technologies; Nucleic Acid Hybridization; Placenta - cytology; Placenta - metabolism; Pregnancy; Thymidine Kinase - genetics
• ISSN: 0740-7750; 1572-9931
• DOI: 10.1007/BF01534722

We describe an in situ hybridization technique which allows rapid identification of species-specific chromosomes in somatic cell hybrid lines. Chromosome preparations from rodent-human hybrid lines are hybridized to biotinylated total human DNA which is subsequently detected by a series of immunocytochemical reactions which culminate in a peroxidase reaction visible by light microscopy. This technique not only allows identification of intact human chromosomes but also fragmented and rearranged human chromosomal segments. We have detected as little as 1 X 10(7) bp of human DNA inserted into a mouse chromosome using this procedure and estimate that the sensitivity of the technique would allow detection of sequences 5- to 10-fold smaller. The usefulness of the technique for screening hybrid cell gene mapping panels is discussed.