Unit

UNIT 24.2 Design, Synthesis, and Amplification of DNA Pools for In Vitro Selection

  1. Bradley Hall1,
  2. John M. Micheletti2,
  3. Pooja Satya2,
  4. Krystal Ogle2,
  5. Jack Pollard3,
  6. Andrew D. Ellington1

Published Online: 1 OCT 2009

DOI: 10.1002/0471142727.mb2402s88

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Hall, B., Micheletti, J. M., Satya, P., Ogle, K., Pollard, J. and Ellington, A. D. 2009. Design, Synthesis, and Amplification of DNA Pools for In Vitro Selection. Current Protocols in Molecular Biology. 88:24.2:24.2.1–24.2.27.

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

    3rd Millennium Corporation, Cambridge, Massachusetts

Publication History

  1. Published Online: 1 OCT 2009
  2. Published Print: OCT 2009

Abstract

Preparation of a random-sequence DNA pool is presented. The degree of randomization and the length of the random sequence are discussed, as is synthesis of the pool using a DNA synthesizer or via commercial synthesis companies. Purification of a single-stranded pool and conversion to a double-stranded pool are presented as step-by-step protocols. Support protocols describe determination of the complexity and skewing of the pool, and optimization of amplification conditions. Curr. Protoc. Mol. Biol. 88:24.2.1-24.2.27. © 2009 by John Wiley & Sons, Inc.

Keywords:

  • In vitro selection;
  • DNA pool synthesis;
  • phosphoramidite DNA synthesis;
  • randomization