Unit

UNIT 5.12 Normalizing cDNA Libraries

  1. Ekaterina A. Bogdanova1,
  2. Irina Shagina2,
  3. Ekaterina V. Barsova1,
  4. Ilya Kelmanson2,
  5. Dmitry A. Shagin1,
  6. Sergey A. Lukyanov1

Published Online: 1 APR 2010

DOI: 10.1002/0471142727.mb0512s90

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Bogdanova, E. A., Shagina, I., Barsova, E. V., Kelmanson, I., Shagin, D. A. and Lukyanov, S. A. 2010. Normalizing cDNA Libraries. Current Protocols in Molecular Biology. 90:V:5.12:5.12.1–5.12.27.

Author Information

  1. 1

    Shemiakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russia

  2. 2

    Evrogen JSC, Moscow, Russia

Publication History

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

Abstract

The characterization of rare messages in cDNA libraries is complicated by the substantial variations that exist in the abundance levels of different transcripts in cells and tissues. The equalization (normalization) of cDNA is a helpful approach for decreasing the prevalence of abundant transcripts, thereby facilitating the assessment of rare transcripts. This unit provides a method for duplex-specific nuclease (DSN)–based normalization, which allows for the fast and reliable equalization of cDNA, thereby facilitating the generation of normalized, full-length-enriched cDNA libraries, and enabling efficient RNA analyses. Curr. Protoc. Mol. Biol. 90:5.12.1-5.12.27. © 2010 by John Wiley & Sons, Inc.

Keywords:

  • cDNA normalization;
  • duplex-specific nuclease (DSN);
  • normalized cDNA library