Unit

UNIT 22.2 Preparation of mRNA for Expression Monitoring

  1. Michael C. Byrne,
  2. Maryann Z. Whitley,
  3. Maximillian T. Follettie

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb2202s49

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Byrne, M. C., Whitley, M. Z. and Follettie, M. T. 2001. Preparation of mRNA for Expression Monitoring. Current Protocols in Molecular Biology. 49:22.2:22.2.1–22.2.13.

Author Information

  1. Genetics Institute, Cambridge, Massachusetts

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JAN 2000

Abstract

The ability to construct comprehensive gene expression profiles comprising hundreds to thousands of genes whose RNA levels are monitored simultaneously represents an exciting new capability in molecular biology. This is accomplished by hybridizing mRNA, which has been quantitatively amplified and labeled with biotin, to DNA chips that display thousands of nucleotides complementary to the mRNAs of interest. In this unit, rationale for starting with poly(A+) vs. total RNA is discussed, and strategies for choosing oligonucleotides for chip design is presented. Protocols on RNA amplification and labeling, and purifying and quantifying the cDNA and in vitro transcription products are included.