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Medizin - Open Access LMU - Teil 09/22 Podcast
Rapid generation of chromosome-specific alphoid DNA probes using the polymerase chain reaction

Rapid generation of chromosome-specific alphoid DNA probes using the polymerase chain reaction

Medizin - Open Access LMU - Teil 09/22 vor 34 Jahren

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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.