Rapid generation of chromosome-specific alphoid DNA probes using the polymerase chain reaction.
Non-isotopic in situ hybridization of chromosome-specific alphoid DNA probes has become a potent tool in the study of numerical aberrations of specific human chromosomes at all stages of the cell cycle. In this paper, we describe approaches for the rapid generation of such probes using the polymerase chain reaction (PCR), and demonstrate their chromosome specificity by fluorescence in situ hybridization to normal human metaphase spreads and interphase nuclei. Oligonucleotide primers for conserved regions of the alpha satellite monomer were used to generate chromosome-specific DNA probes from somatic hybrid cells containing various human chromosomes, and from DNA libraries from sorted human chromosomes. Oligonucleotide primers for chromosome-specific regions of the alpha satellite monomer were used to generate specific DNA probes for the pericentromeric heterochromatin of human chromosomes 1, 6, 7, 17 and X directly from human genomic DNA.
Dunham
I
I
Department of Genetics, Howard Hughes Medical Institute, Washington University School of Medicine, St. Louis, MO 63110.
Lengauer
C
C
Cremer
T
T
Featherstone
T
T
D000818:Animals; D001483:Base Sequence; D002453:Cell Cycle; D002460:Cell Line; D002869:Chromosome Aberrations; D002877:Chromosomes, Human; D002878:Chromosomes, Human, Pair 1; D002886:Chromosomes, Human, Pair 17; D002896:Chromosomes, Human, Pair 6; D006224:Cricetinae; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D015342:DNA Probes; D006570:Heterochromatin / Q000502:physiology* / Q000648:ultrastructure; D006801:Humans; D008677:Metaphase; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction / Q000379:methods*; D014960:X Chromosome
D015342:DNA Probes; D006570:Heterochromatin; D009838:Oligodeoxyribonucleotides; D004247:DNA