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J Mol Biol. 2009 Jan 23;385(3):714-7. doi: 10.1016/j.jmb.2008.11.035. Epub 2008 Nov 27.

Characterization of DNA-binding proteins using multiplexed competitor EMSA.

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  • 1Center for Cardiovascular Genetics, Department of Medicine, University College London, 5 University Street, London WC1E 6JF, UK. Andrew.J.P.Smith@ucl.ac.uk.

Abstract

We describe a low-cost high-throughput technique to characterize nuclear protein DNA-binding interactions. This technique, known as Multiplexed Competitor Electrophoretic Mobility Shift Assay, uses a series of multiplexed oligonucleotide DNA consensus competitors, in combination with a standard electrophoretic mobility shift assay procedure, to efficiently characterize DNA-binding proteins. We show utility for the method to identify a previously unreported hepatocyte nuclear factor-3 site created in intron 8 of the lipoprotein lipase gene by a common single-nucleotide polymorphism (rs327).

PMID:
19059416
[PubMed - indexed for MEDLINE]
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