RNA Detection

3.4 Imaging RNA in
Live Cells by Using
SPAAC Reaction

1. Culture HeLa cells in complete DMEM supplemented with
   10% FBS (seeNote 1).


2. Seed 0.3–0.4 million HeLa cells in a 35 mm dish having a glass
   surface. Place the dish in the CO 2 incubator at 37C for nearly
   12 h (seeNote 13).


3. Remove the media from the culture dish and carefully transfer
   600 μL of the transfection mix containing AMUTP into the
   dish.

Fig. 3Imaging cellular RNA transcription using AMUTP. Cultured HeLa cells were

transfected with AMUTP (1 mM) for 1 h using DOTAP. The cells were then fixed,

permeabilized and the AMU-labeling was detected by performing CuAAC

reaction with Alexa 594 alkyne. This figure has been reproduced by

permission of Nucleic Acids Research: Oxford Journals [24]

366 Anupam A. Sawant et al.