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An Optimized Protocol for Electrophoretic Mobility Shift Assay Using Infrared Fluorescent Dye-labeled Oligonucleotides
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Biochemistry
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JoVE Journal Biochemistry
An Optimized Protocol for Electrophoretic Mobility Shift Assay Using Infrared Fluorescent Dye-labeled Oligonucleotides

An Optimized Protocol for Electrophoretic Mobility Shift Assay Using Infrared Fluorescent Dye-labeled Oligonucleotides

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15,080 Views

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09:58 min

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November 29, 2016

DOI:

10.3791/54863-v

DOI:
10.3791/54863-v

•

09:58 min
November 29, 2016

•

5 Views
, ,
1Department of Biological Sciences,University of Illinois at Chicago

Summary

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We describe here an optimized protocol of fluorescent Electrophoretic Mobility Shift Assays (fEMSA) using purified SOX-2 proteins together with infrared fluorescent dye-labeled DNA probes as a case study to tackle an important biological question.

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Electrophoretic Mobility Shift AssayInfrared Fluorescent DyeOligonucleotidesProtein-DNA InteractionsMolecular BiologyBiochemistryNon-radioactive ProbesNative Polyacrylamide GelTBE BufferGlycerolAnnealingDouble-stranded DNA ProbesKlenow FragmentEDTAPCRUnlabeled Probes

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Hsieh, Y., Alqadah, A., Chuang, C. An Optimized Protocol for Electrophoretic Mobility Shift Assay Using Infrared Fluorescent Dye-labeled Oligonucleotides. J. Vis. Exp. (117), e54863, doi:10.3791/54863 (2016).

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