Human Genetics

, Volume 77, Issue 4, pp 366–370

Use of whole cosmid cloned genomic sequences for chromosomal localization by non-radioactive in situ hybridization

  • J. E. Landegent
  • N. Jansen in de Wal
  • R. W. Dirks
  • M. van der Ploeg
Original Investigations

DOI: 10.1007/BF00291428

Cite this article as:
Landegent, J.E., Jansen in de Wal, N., Dirks, R.W. et al. Hum Genet (1987) 77: 366. doi:10.1007/BF00291428

Summary

We report a general procedure which allows the application of whole cosmid cloned genomic sequences for non-radioactive in situ hybridization. The presence of highly repetitive sequences, like Alu and Kpn fragments, is eliminated through competition hybridization with Cot-1 DNA. The method has been tested and optimized with several randomly chosen cosmids of the human thyroglobulin (Tg) gene (8q24). At present, the procedure can be performed with three of the four tested individual cosmids. In cases where a single clone does not result in a specific signal, a larger fragment may be required, which can be accomplished by using two (partially overlapping) cosmids of the same region. The advantages and further potentialities of such a hybridization approach are discussed.

Copyright information

© Springer-Verlag 1987

Authors and Affiliations

  • J. E. Landegent
    • 1
  • N. Jansen in de Wal
    • 1
  • R. W. Dirks
    • 1
    • 2
  • M. van der Ploeg
    • 1
  1. 1.Department of Cytochemistry and Cytometry, Sylvius LaboratoriesUniversity of LeidenLeidenThe Netherlands
  2. 2.Division of Molecular BiologyThe Netherlands Cancer Institute (Antonie van Leeuwenhoekhuis)AmsterdamThe Netherlands
  3. 3.Biomedical Sciences DivisionLawrence Livermore National LaboratoryLivermoreUSA
  4. 4.Department of Biochemistry and Molecular BiologyHarvard UniversityCambridgeUSA