A protocol for preparing, characterizing and using three RNA-specific, live cell imaging probes: E36, E144 and F22.
This protocol outlines a methodology for the preparation and characterization of three RNA-specific fluorescent probes (E36, E144 and F22) and their use in live cell imaging. It describes a detailed procedure for their chemical synthesis and purification; serial product characterization and quality control tests, including measurements of their fluorescence properties in solution, measurement of RNA specificity and analysis of cellular toxicity; and live cell staining and counterstaining with Hoechst or DAPI. Preparation and application of these RNA imaging probes takes 1 week.
Li
Qian
Q
Department of Chemistry, New York University, New York, NY 10003, USA.
Chang
Young-Tae
YT
D005456:Fluorescent Dyes / Q000138:chemical synthesis* / Q000737:chemistry / Q000302:isolation & purification; D015336:Molecular Probe Techniques; D012313:RNA / Q000737:chemistry*
D005456:Fluorescent Dyes; D012313:RNA