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Papers In Press, published online ahead of print May 1, 2008
J. Lipid Res., doi:10.1194/jlr.D800002-JLR200

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Journal of Lipid Research, Vol. 49, 1126-1129, May 2008
Copyright © 2008 by American Society for Biochemistry and Molecular Biology

Methods

A rapid, microplate SNP genotype assay for the leptin^ob allele

Angie T. Oler and Alan D. Attie¹

Department of Biochemistry, University of Wisconsin-Madison, Madison, WI 53706

The online version of this article (available at http://www.jlr.org) contains supplementary data.

Published, JLR Papers in Press, February 13, 2008.

¹ To whom correspondence should be addressed. e-mail: attie{at}biochem.wisc.edu

A nonsense mutation in the mouse leptin gene causes genetic obesity. As a result of extensive research in the field of obesity, the use of leptin^ob mice is widespread. This mutation renders mice sterile, creating the need to breed heterozygous mice. For this reason, leptin^ob genotyping is necessary. To date, gel-based assays have been used for genotyping. Using the Invader Plus® assay for single nucleotide polymorphism (SNP) detection, we have developed a gel-free microplate SNP assay for genotyping leptin^wt and leptin^ob alleles.

Supplementary key words obesity • obese • single nucleotide polymorphism

Abbreviations: FAM, fluorescein; FRET, fluorescence resonant energy transfer; RFU, relative fluorescence units; SNP, single nucleotide polymorphism

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