Unit

UNIT 9.2 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 DEC 2009

DOI: 10.1002/0471142700.nc0902s39

Current Protocols in Nucleic Acid Chemistry

Current Protocols in Nucleic Acid Chemistry

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 Nucleic Acid Chemistry. 39:9.2:9.2.1–9.2.28.

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 DEC 2009
  2. Published Print: DEC 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. Nucleic Acid Chem. 39:9.2.1-9.2.28. © 2009 by John Wiley & Sons, Inc.

Keywords:

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