Unit

UNIT 4.26 Preparation of mRNA for Expression Monitoring

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

Published Online: 1 NOV 2001

DOI: 10.1002/0471142301.ns0426s16

Current Protocols in Neuroscience

Current Protocols in Neuroscience

How to Cite

Byrne, M. C., Whitley, M. Z. and Follettie, M. T. 2001. Preparation of mRNA for Expression Monitoring. Current Protocols in Neuroscience. 16:4.26:4.26.1–4.26.13.

Author Information

  1. Genetics Institute/Wyeth Research, Cambridge, Massachusetts

Publication History

  1. Published Online: 1 NOV 2001
  2. Published Print: AUG 2001

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+) versus 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.