Unit

UNIT 9.3 In Vitro Selection of RNA Aptamers to a Protein Target by Filter Immobilization

  1. Bradley Hall1,
  2. Seyed Arshad2,
  3. Kyunghyun Seo2,
  4. Catherine Bowman2,
  5. Meredith Corley2,
  6. Sulay D. Jhaveri3,
  7. Andrew D. Ellington1,2

Published Online: 1 MAR 2010

DOI: 10.1002/0471142700.nc0903s40

Current Protocols in Nucleic Acid Chemistry

Current Protocols in Nucleic Acid Chemistry

How to Cite

Hall, B., Arshad, S., Seo, K., Bowman, C., Corley, M., Jhaveri, S. D. and Ellington, A. D. 2010. In Vitro Selection of RNA Aptamers to a Protein Target by Filter Immobilization. Current Protocols in Nucleic Acid Chemistry. 40:9.3:9.3.1–9.3.27.

Author Information

  1. 1

    Department of Chemistry and Biochemistry, University of Texas, Austin, Texas

  2. 2

    Freshman Research Initiative, University of Texas, Austin, Texas

  3. 3

    Nova Research, Inc., Alexandria, Virginia

Publication History

  1. Published Online: 1 MAR 2010
  2. Published Print: MAR 2010

Abstract

This unit describes the selection of aptamers from a pool of single-stranded RNA by binding to a protein target. Aptamers generated from this selection experiment can potentially act as protein function inhibitors, and may find applications as therapeutic or diagnostic reagents. A pool of dsDNA is used to generate an ssRNA pool, which is mixed with the protein target. Bound complexes are separated from unbound reagents by filtration, and the RNA:protein complexes are amplified by a combination of reverse transcription, PCR, and in vitro transcription. Curr. Protoc. Nucleic Acid Chem. 40:9.3.1-9.3.27. © 2010 by John Wiley & Sons, Inc.

Keywords:

  • aptamer;
  • in vitro selection;
  • affinity reagent;
  • filter binding assay;
  • SELEX