Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (230K) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Lopez-Crapez, E.
Right arrow Articles by Mathis, G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Lopez-Crapez, E.
Right arrow Articles by Mathis, G.
Related Collections
Right arrow Polymorphism/mutation detection
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 2001, Vol. 29, No. 14 e70
© 2001 Oxford University Press

A homogeneous europium cryptate-based assay for the diagnosis of mutations by time-resolved fluorescence resonance energy transfer

E. Lopez-Crapez*, H. Bazin1, E. Andre, J. Noletti, J. Grenier and G. Mathis1

Centre de Recherche en Cancérologie, C.R.L.C. Val d’Aurelle, Parc Euromédecine, 34298 Montpellier Cedex 5, France and 1CIS Bio International, Etablissement de Marcoule, R&D/HTRF, BP 84 175, 30 204 Bagnols/Cèze Cedex, France

Oligonucleotide ligation assay (OLA) is considered to be a very useful methodology for the detection and characterization of mutations, particularly for clinical purposes. The fluorescence resonance energy transfer between a fluorescent donor and a suitable fluorophore as acceptor has been applied in the past to several scientific fields. This technique is well adapted to nucleic acid analysis such as DNA sequencing, DNA hybridization and polymerase chain reaction. We describe here a homogeneous format based on the use of a rare earth cryptate label as donor: tris-bipyridine-Eu3+. The long-lived fluorescence of this label makes it possible to reach a high sensitivity by using a time-resolved detection mode. A non-radiative energy transfer technology, known as time-resolved amplification of cryptate emission (TRACE®) characterized by a temporal and spectral selectivity has been developed. The TRACE® detection of characterized single nucleotide polymorphism using the OLA for allelic discrimination is proposed. We demonstrate the potentialities of this OLA–TRACE® methodology through the analysis of K-ras oncogene point mutations.

* To whom correspondence should be addressed. Tel: +33 4 67 61 30 48; Fax: +33 4 67 63 28 73; Email: ecrapez@valdorel.fnclcc.fr


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.