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In Vitro Selection of ssDNA Aptamers Using Biotinylated Target Proteins

  • Günter Mayer
  • Thomas Höver
Protocol
Part of the Methods in Molecular Biology™ book series (MIMB, volume 535)

Abstract

Aptamers are single-stranded nucleic acids that bind specifically to a target molecule and thus often inhibit target-associated biological functions. Aptamers have been described for a series of target molecules including peptides, proteins, and even living cells. Besides RNA and 2′-modified RNA molecules also ssDNA molecules can be subjected to in vitro selection protocols aiming at the enrichment of ssDNA aptamers. ssDNA aptamers can be selected using the SELEX procedure (systematic enrichment of ligands by exponential amplification) from libraries of randomized single-stranded DNA with a diversity of up to 1016 different molecules. In repetitive selection cycles, the library is incubated with the target of choice and separation of non-binding sequences from bound sequences is achieved by distinct separation methods. The bound molecules are specifically eluted and amplified, thus representing the starting library for the next cycle. Thereby, an enriched population of aptamers is evolved. Here we describe a generalized in vitro selection experiment aiming at the enrichment of ssDNA aptamers using biotinylated target molecules. This procedure allows the application of streptavidin–biotin chemistry to separate bound from unbound DNA species during the selection process.

Key words

SELEX aptamers ssDNA in vitro selection biotin streptavidin 

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Günter Mayer
    • 1
  • Thomas Höver
    • 1
  1. 1.Life and Medical Sciences, Program Unit Chemical Biology and Medicinal ChemistryUniversity of BonnBonnGermany

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