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Quantitative Analysis of MicroRNA in Blood Serum with Protein-Facilitated Affinity Capillary Electrophoresis

  • Maxim V. Berezovski
  • Nasrin Khan
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1039)

Abstract

MicroRNAs play an important role in gene regulation and disease etiology and are blood-based biomarkers of diseases. Here, we describe a protein-facilitated affinity capillary electrophoresis (ProFACE) method for ultra-sensitive direct miRNA detection as low as 300,000 molecules in 1 mL of blood serum, using single-stranded DNA binding protein (SSB) and double-stranded RNA binding protein (p19) as separation enhancers. This method utilizes either the selective binding of SSB to a fluorescent single-stranded DNA/RNA probe or the binding of p19 to miRNA–RNA probe duplex.

Key words

MicroRNA SSB p19 protein Blood serum Capillary electrophoresis Laser-induced fluorescence detection 

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Copyright information

© Springer Science+Business Media, New York 2013

Authors and Affiliations

  • Maxim V. Berezovski
    • 1
  • Nasrin Khan
    • 2
  1. 1.University of OttawaOttawaCanada
  2. 2.Department of ChemistryUniversity of OttawaOttawaCanada

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