Unit

UNIT 9.6 In Vitro Selection Using Modified or Unnatural Nucleotides

  1. Gwendolyn M. Stovall1,
  2. Robert S. Bedenbaugh1,
  3. Shruti Singh1,
  4. Adam J. Meyer1,
  5. Paul J. Hatala2,
  6. Andrew D. Ellington1,
  7. Bradley Hall2

Published Online: 26 MAR 2014

DOI: 10.1002/0471142700.nc0906s56

Current Protocols in Nucleic Acid Chemistry

Current Protocols in Nucleic Acid Chemistry

How to Cite

Stovall, G. M., Bedenbaugh, R. S., Singh, S., Meyer, A. J., Hatala, P. J., Ellington, A. D. and Hall, B. 2014. In Vitro Selection Using Modified or Unnatural Nucleotides. Current Protocols in Nucleic Acid Chemistry. 9:9.6:9.6.1–9.6.33.

Author Information

  1. 1

    The University of Texas at Austin, Austin, Texas

  2. 2

    Altermune Technologies LLC, Austin, Texas

Publication History

  1. Published Online: 26 MAR 2014

Abstract

Incorporation of modified nucleotides into in vitro RNA or DNA selections offers many potential advantages, such as the increased stability of selected nucleic acids against nuclease degradation, improved affinities, expanded chemical functionality, and increased library diversity. This unit provides useful information and protocols for in vitro selection using modified nucleotides. It includes a discussion of when to use modified nucleotides; protocols for evaluating and optimizing transcription reactions, as well as confirming the incorporation of the modified nucleotides; protocols for evaluating modified nucleotide transcripts as template in reverse transcription reactions; protocols for the evaluation of the fidelity of modified nucleotides in the replication and the regeneration of the pool; and a protocol to compare modified nucleotide pools and selection conditions. Curr. Protoc. Nucleic Acid Chem. 56:9.6.1-9.6.33. © 2014 by John Wiley & Sons, Inc.

Keywords:

  • in vitro selection;
  • aptamer;
  • ribozymes;
  • deoxyribozymes;
  • modified nucleotides;
  • unnatural nucleotides