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Multicolor fluorescence in situ hybridization on metaphase chromosomes and interphase Halo-preparations using cosmid and YAC clones for the simultaneous high resolution mapping of deletions in the dystrophin gene

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Abstract

We report on multicolor fluorescence in situ hybridization protocols for the simultaneous visualization of deletion-prone regions for carrier detection of Duchenne/ Becker (DMD/BMD) muscular dystrophy. Cosmid and yeast artificial chromosome (YAC) clones specific for preferentially deleted subregions of the dystrophin gene were labeled differentially and detected with three different fluorochromes using digital imaging microscopy. This approach allows for an assessment of the carrier status of female relatives even in families where no index patient is available. Cosmid and YAC clones, and different probe-generation protocols are compared with respect to their feasibility for carrier detection. The use of histone-depleted interphase nuclei (Halo-preparations) for deletion mapping is demonstrated and shown to have a resolution power of 5 kb.

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Authors and Affiliations

  1. Institut für Humangenetik und Anthropologie der Universität, Im Neuenheimer Feld 328, D-69120, Heidelberg, Germany

    Chintana Tocharoentanaphol, Marion Cremer, Evelin Schröck, Karin Kilian, Thomas Cremer & Thomas Ried

  2. Faculty of Medicine, Department of Genetics, Wassenaarseweg 72, 2300, RA Leiden, The Netherlands

    Lau Blonden

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  1. Chintana Tocharoentanaphol
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  2. Marion Cremer
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  3. Evelin Schröck
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  4. Lau Blonden
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  5. Karin Kilian
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  6. Thomas Cremer
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  7. Thomas Ried
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Tocharoentanaphol, C., Cremer, M., Schröck, E. et al. Multicolor fluorescence in situ hybridization on metaphase chromosomes and interphase Halo-preparations using cosmid and YAC clones for the simultaneous high resolution mapping of deletions in the dystrophin gene. Hum Genet 93, 229–235 (1994). https://doi.org/10.1007/BF00212014

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  • DOI: https://doi.org/10.1007/BF00212014

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