Advertisement

Oligonucleotide PRINS DNA Synthesis

  • John R. Gosden
  • Diane Lawson
Protocol
  • 85 Downloads
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

The technique for labeling chromosomes by annealing an oligonucleotide DNA primer to the denatured DNA of chromosome preparations on glass slides and extending it enzymatically in situ with the incorporation of labeled nucleotides was first described by Koch et al. in 1989 (1). Since then, the technique has been greatly improved in reliability, sensitivity, and resolution, and now provides a viable, rapid alternative to conventional fluorescence in situ hybridization (FISH) for many investigations, particularly the identification of chromosome aneuploidy in metastatic tissues and antenatal diagnosis and the analysis of the human chromosome complement of somatic hybrid cell lines (2, 3, 4, 5, 6).

Keywords

Amersham Pharmacia Biotech Wash Buffer Skim Milk Powder Chromosome Aneuploidy Stop Buffer 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Koch, J. E., Kølvraa, S., Petersen, K. B., Gregersen, N., and Bolund, I. (1989) Oligo-nucleotide-priming methods for the chromosome-specific labelling of alpha satellite DNA in situ. Chromosoma 98, 259–265.PubMedCrossRefGoogle Scholar
  2. 2.
    Gosden, J., Hanratty, D., Starling, J., Fantes, J., Mitchell, A., and Porteous, D. (1991) Oli-gonucleotide primed in situ DNA synthesis (PRINS): a method for chromosome mapping, banding and investigation of sequence organization. Cytogenet. Cell Genet. 57, 100–104.PubMedCrossRefGoogle Scholar
  3. 3.
    Gosden, J. and Lawson, D. (1994) Rapid chromosome identification by oligonucleotide primed in situ DNA synthesis (PRINS). Hum. Mol. Genet. 3, 931–946.PubMedCrossRefGoogle Scholar
  4. 4.
    Gosden, J. and Lawson, D. (1995) Instant PRINS: a rapid method for chromosome identification by detecting repeated sequences in situ. Cytogenet. Cell Genet. 68, 57–60.PubMedCrossRefGoogle Scholar
  5. 5.
    Hindkjaer, J., Koch, J., Terkelsen, C., Brandt, C. A., Kølvraa, S., and Bolund, L. (1994) Fast, sensitive multicolour detection of nucleic acids by primed in situ labelling (PRINS). Cytogenet. Cell Genet. 66, 152–154.PubMedCrossRefGoogle Scholar
  6. 6.
    Speel, E. J. M., Lawson, D., Hopman, A. H. N., and Gosden, J. (1995) Multi-PRINS: multiple sequential oligonucleotide primed in situ DNA synthesis reactions label specific chromosomes and produce bands. Hum. Genet. 95, 29–33.PubMedCrossRefGoogle Scholar
  7. 7.
    Spowart, G. (1994) Mitotic metaphase chromosome preparation from peripheral blood for high resolution, in Methods in Molecular Biology, vol. 29: Chromosome Analysis Protocols (Gosden J. R., ed.), Humana, Totowa, NJ, pp. 1–10.CrossRefGoogle Scholar
  8. 8.
    Fletcher, J. (1994) Immortalized cells lines: chromosome preparation and binding, in Methods in Molecular Biology, vol. 29: Chromosome Analysis Protocols (Gosden, J. R., ed.), Humana, Totowa, NJ, pp. 51–57.CrossRefGoogle Scholar
  9. 9.
    Speel, E. J. M., Lawson, D., Ramachers, F. C. S., Gosden, J. R., and Hopman, A. H. N. (1997) Combined immunocytochemistry and PRINS DNA synthesis for simultaneous detection of phenotypic and genomic parameters in cells, in Methods in Molecular Biology, vol. 71, PRINS and In Situ PCR Protocols (Gosden, J. R., ed.), Humana, Totowa, NJ, pp. 53–59.Google Scholar
  10. 10.
    Multiple sequential oligonucleotide primed in situ DNA synthesis (MULTI-PRINS) (1997), in Methods in Molecular Biology, vol. 71, PRINS and In Situ PCR Protocols, (Gosden, J. R., ed.), Humana, Totowa, NJ, pp. 39–44.Google Scholar
  11. 11.
    Warburton, P. E., Haaf, T., Gosden, J., Lawson, D., and Willard, H. F. (1996) Characterization of a chromosome-specific chimpanzee alpha satellite subset: evolutionary relationship to subsets on human chromosomes. Genomics 33, 220–228.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2000

Authors and Affiliations

  • John R. Gosden
    • 1
  • Diane Lawson
    • 1
  1. 1.MRC Human Genetics UnitWestern General HospitalEdinburghUK

Personalised recommendations