UNIT 22.2 Preparation of mRNA for Expression Monitoring
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
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.
- Published Online: 1 MAY 2001
- Published Print: JAN 2000
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.