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 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.
D000818:Animals; D001853:Bone Marrow / Q000201:enzymology; D001854:Bone Marrow Cells; D002460:Cell Line; D002875:Chromosomes / Q000648:ultrastructure*; D006224:Cricetinae; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D005260:Female; D006801:Humans; D006822:Hybrid Cells / Q000166:cytology* / Q000201:enzymology; D007668:Kidney; D009693:Nucleic Acid Hybridization; D010920:Placenta / Q000166:cytology / Q000378:metabolism; D011247:Pregnancy; D013937:Thymidine Kinase / Q000235:genetics
D004247:DNA; D013937:Thymidine Kinase