UNIT 7.12 Flow Cytometric Analysis of RNA Synthesis by Detection of Bromouridine Incorporation
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Cytometry
How to Cite
Larsen, J. K., Jensen, P. Ø. and Larsen, J. 2001. Flow Cytometric Analysis of RNA Synthesis by Detection of Bromouridine Incorporation. Current Protocols in Cytometry. 12:7.12:7.12.1–7.12.11.
- Published Online: 1 MAY 2001
- Published Print: APR 2000
RNA synthesis has traditionally been investigated by a laborious and time-consuming radiographic method involving incorporation of tritiated uridine. Now a faster non-radioactive alternative has emerged, based on immunocytochemical detection. This method utilizes the brominated RNA precursor bromouridine, which is taken into a cell, phosphorylated, and incorporated into nascent RNA. The BrU-substituted RNA is detected by permeabilizing the cells and staining with certain anti-BrdU antibodies. This dynamic approach yields information complementing that provided by cellular RNA content analysis at a given time and may be of value in studies of cellular activation and gene expression.