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Nucleic Acids Research Advance Access published online on December 12, 2006
Nucleic Acids Research, doi:10.1093/nar/gkl1046
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© 2006 The Author(s).
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Methods Online

A semi-automated high-throughput approach to the generation of transposon insertion mutants in the nematode Caenorhabditis elegans

Yohann Duverger1,2,3, Jérôme Belougne4, Sarah Scaglione1,2,3, Dominique Brandli4, Christophe Beclin4 and Jonathan J. Ewbank1,2,3,*

1 Centre d'Immunologie de Marseille-Luminy, Université de la Méditerranée Case 906, 13288 Marseille cedex 9, France 2 INSERM U631, 13288 Marseille, France 3 CNRS UMR6102, 13288 Marseille, France 4 CNRS, Institut de Biologie du Développement de Marseille-Luminy Marseille, France

*To whom correspondence should be addressed. Tel: +33 491 269 472; Fax: +33 491 269 430; Email: ewbank{at}ciml.univ-mrs.fr

Received September 5, 2006. Revised October 18, 2006. Accepted November 3, 2006.

The generation of a large collection of defined transposon insertion mutants is of general interest to the Caenorhabditis elegans research community and has been supported by the European Union. We describe here a semi-automated high-throughput method for mutant production and screening, using the heterologous transposon Mos1. The procedure allows routine culture of several thousand independent nematode strains in parallel for multiple generations before stereotyped molecular analyses. Using this method, we have already generated >17 500 individual strains carrying Mos1 insertions. It could be easily adapted to forward and reverse genetic screens and may influence researchers faced with making a choice of model organism.


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