Detection of species specific chromosomes in somatic cell hybrids

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 immunocyt...

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Bibliographic Details
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
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Summary: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.
ISSN:0740-7750
1572-9931
DOI:10.1007/BF01534722