Using a novel approach — recombineering — to generate odf2 null alleles

Abstract

This article uses a real-life example to illustrate the concept and methodology of recombineering, a revolutionary genetic engineering technique based on phage-mediated homologous recombination. A step-by-step approach is presented along with a flow diagram, from obtaining gene-harboring BACs to the in vitro generation of a conditional null allele. This method can be used to target any gene at any position, without the knowledge or use of any restriction site. The extensive applicability of recombineering to gene manipulation is discussed.

Keywords

gene manipulation / recombineering / odf2

References

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Acknowledgments

This work was carried out in the laboratory of Dr. Jonathan Eggenschwiler at the Department of Molecular Biology at Princeton University, whose expertise and support continue to be invaluable and are much appreciated.

RIGHTS & PERMISSIONS

2014 Higher Education Press and Springer-Verlag Berlin Heidelberg