- A protocol for preparing, characterizing and using three RNA-specific, live cell imaging probes: E36, E144 and F22.
A protocol for preparing, characterizing and using three RNA-specific, live cell imaging probes: E36, E144 and F22.
Nature protocols (2007-04-05)
Qian Li, Young-Tae Chang
PMID17406504
ABSTRACT
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.
MATERIALS
Product Number
Brand
Product Description
Sigma-Aldrich
4′,6-Diamidino-2-phenylindole dihydrochloride, powder, BioReagent, suitable for cell culture, ≥98% (HPLC and TLC), suitable for fluorescence
Sigma-Aldrich
Deoxyribonuclease I from bovine pancreas, Standardized vial containing 2,000 Kunitz units of DNase I (D4527), vial of ≥0.25 mg total protein