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

DOI:
10.3791/54863-v

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09:58 min

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November 29, 2016

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

Chapitres

  • 00:05Titre
  • 00:32Preparation of Gel, Infrared Fluorescent Dye-labeled Probes and Unlabeled Probes
  • 04:49Binding Reaction and Electrophoresis
  • 06:37Imaging
  • 07:35Results: Electrophoretic Mobility Shift Assays Using Purified Proteins and Dye-labeled DNA Probes
  • 08:57Conclusion

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.

Tags

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

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