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In situ method for differentiating homologous DNA sequences, useful in medical diagnosis and research, by fluorescence microscopic comparison of chromosomes or interphase nuclei
In situ method for differentiating homologous DNA sequences, useful in medical diagnosis and research, by fluorescence microscopic comparison of chromosomes or interphase nuclei
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机译：通过染色体或相间核的荧光显微镜比较，用于区分同源DNA序列的原位方法，可用于医学诊断和研究
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摘要
In situ method for differentiating homologous DNA sequences in which chromosomes or interphase nuclei of the test material are evaluated and compared, by fluorescence microscopy, for presence of polymorphic differences at the DNA level. In situ method for differentiating homologous DNA sequences in which chromosomes or interphase nuclei of the test material are evaluated and compared, by fluorescence microscopy (FM), for presence of polymorphic differences at the DNA level. These DNA sequences (or segments of them) are examined for known DNA polymorphisms (P), then detected (P) that are relevant to the particular analysis being done, are used to select, amplify and label (e.g. with fluorescent dyes or haptens) one or more DNA sequences or bacterial artificial chromosomes (BAC). These materials are combined, e.g. in chromosome-specific sets and, together with preferably Cot-1 DNA, are denatured to produce single-stranded probes, containing sequences homologous with the material being examined, and these are hybridized to carriers, preferably at about 37[deg]C, then washed, rinsed, dehydrated, dried and counterstained (e.g. with DAPI) for evaluation by FM. The fluorescent signals from the test material, and those for the carrier-hybridized DNA probes, are compared and evaluated for differences in DNA sequences as regards reproducible intensity differences.
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