Digoxigenated oligonucleotide probes specific for simple repeats in DNA fingerprinting and hybridization in situ (original) (raw)

Summary

A fast, reproducible and non-hazardous technique for non-isotopic DNA fingerprinting is presented. The method is based on digoxigenated oligonucleotides, which are specific for simple repetitive DNA sequences. The use of digoxigenin/ anti-digoxigenin detection avoids many drawbacks inherent in e.g. the biotin/streptavidin system which often causes a poor signal-to-background ratio. Synthesis and purification of digoxigenated oligonucleotides and their use in filter hybridization are described in detail. Hybridization patterns obtained with four different radioactively labeled oligonucleotides have been compared with those of the respective digoxigenated probes. When slightly less stringent hybridization conditions are applied for digoxigenated oligonucleotides than for those labeled with 32P, the signal intensities are satisfying but additional minor bands occur as a result of the reduced strigency. With one explainable exception, these bands increase the information content of the fingerprint. In addition, hybridization of the digoxigenated (CAC)5 probe has been performed in situ with human metaphase chromosomes. The hybridization patterns in many mitoses resemble R-bands.

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Authors and Affiliations

1. Max-Planck-Institut für Psychiatrie, Am Klopferspitz 18A, D-8033, Martinsried, Federal Republic of Germany
   Hans Zischler, Renate Schäfer & Jörg T. Epplen
2. Institut für Humangenetik der Universität, Koellikerstrasse 2 A, D-8700, Würzburg, Federal Republic of Germany
   Indrajit Nanda & Michael Schmid

Authors

1. Hans Zischler
2. Indrajit Nanda
3. Renate Schäfer
4. Michael Schmid
5. Jörg T. Epplen

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Zischler, H., Nanda, I., Schäfer, R. et al. Digoxigenated oligonucleotide probes specific for simple repeats in DNA fingerprinting and hybridization in situ.Hum Genet 82, 227–233 (1989). https://doi.org/10.1007/BF00291160

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• Received: 20 January 1989
• Revised: 08 February 1989
• Issue date: June 1989
• DOI: https://doi.org/10.1007/BF00291160

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