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PNA-FISH Technique for In Situ Assessment of Aneuploidy

  • Franck PellestorEmail author
  • Petra Paulasova
  • Samir Hamamah
Protocol
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Part of the Springer Protocols Handbooks book series (SPH)

Peptide nucleic acid in situ labeling (PNA-FISH) techniques provide an attractive alternative to conventional FISH and PRINS procedures for chromosomal in situ detection. PNA probes are synthetic DNA analogs with uncharged polyamide backbones. The PNA labeling reaction presents several advantages (specificity, rapidity, discriminating ability, short length of the PNA probes) that make it very attractive for cytogenetic purposes. The efficiency of centromeric and telomeric PNA probes has recently been demonstrated on human cell preparations. Their adaptation for human cells has allowed the development of new and fast molecular cytogenetic procedures, which can advantageously be used to complement FISH and PRINS techniques for the in situ assessment of aneuploidy.

Keywords

Peptide Nucleic Acid Propidium Iodide Solution Rubber Cement Deoxyribose Phosphate Base Pairing Rule 
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.

Notes

Acknowledgments

The experiments performed to elaborate this protocol were partially supported by Ferring Pharmaceuticals and Organon France. P. Paulasova was supported by Czech fellowship VZ FNM 00000064203.

References

  1. Lansdorp PM, Verwoerd NP, van de Rijke FM, Dragowska V, Little MT, Dirks RW, Raap AK, Tanke HJ (1996) Heterogeneity in telomere length of human chromosomes. Hum Mol Genet 5:685–691CrossRefPubMedGoogle Scholar
  2. Nielsen PE (2001) Peptide nucleic acid: A versatile tool in genetic diagnostics and molecular biology. Curr Opin Biotechnol 12:16–20CrossRefPubMedGoogle Scholar
  3. Nielsen PE, Egholm M (1999) An introduction to peptide nucleic acid. Curr Issues Mol Bio 1:89–104Google Scholar
  4. Nielsen PE, Egholm M, Berg RH, Buchardt O (1991) Sequence-selective recognition of DNA by strand displacement with a thymine-substituted polyamide. Science 254:1497–1500CrossRefPubMedGoogle Scholar
  5. Paulasova P, Andräo B, Diblik J, Macek M, Pellestor F (2004) The peptide nucleic acids as probes for chromosomal analysis: Application to human oocytes, polar bodies and preimplantation embryos. Mol Hum Reprod 10:467–472CrossRefPubMedGoogle Scholar
  6. Pellestor F, Andräo B, Taneja K, Williams B (2003) PNA on human sperm: A new approach for in situ aneuploidy estimation. Eur J Hum Genet 11:337–341CrossRefPubMedGoogle Scholar
  7. Ray A, Norden B (2000) Peptide nucleic acid (PNA): Its medical and biotechnical applications and promise for the future. FASEB J 14:1041–1060PubMedGoogle Scholar
  8. Taneja KL, Chavez EA, Coull J, Lansdorp PM (2001) Multicolor fluorescence in situ hybridization with peptide nucleic acid probes for enumeration of specific chromosomes in human cells. Genes Chromosomes Cancer 30:57–63CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2009

Authors and Affiliations

  • Franck Pellestor
    • 1
    Email author
  • Petra Paulasova
    • 2
  • Samir Hamamah
    • 1
  1. 1.Service de Médecine et Biologie de la Reproduction, CHRU Montpellier, Hopital Arnaud de VilleneuveINSERM U847Montpellier Cedex 5France
  2. 2.Centre of Assisted Reproduction and Reproductive GeneticsPrague 5Czech Republic

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