Unit

UNIT 7.12 Flow Cytometric Analysis of RNA Synthesis by Detection of Bromouridine Incorporation

  1. Jørgen K. Larsen,
  2. Peter Østrup Jensen,
  3. Jacob Larsen

Published Online: 1 MAY 2001

DOI: 10.1002/0471142956.cy0712s12

Current Protocols in Cytometry

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.

Author Information

  1. The Finsen Laboratory, Rigshospitalet, Copenhagen, Denmark

Publication History

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

Abstract

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.