A protocol for preparing, characterizing and using three RNA-specific, live cell imaging probes: E36, E144 and F22

Qian Li; Young-Tae Chang

doi:10.1038/nprot.2006.484

A protocol for preparing, characterizing and using three RNA-specific, live cell imaging probes: E36, E144 and F22

Nat Protoc. 2006;1(6):2922-32. doi: 10.1038/nprot.2006.484.

Authors

Qian Li¹, Young-Tae Chang

Affiliation

¹ Department of Chemistry, New York University, New York, NY 10003, USA.

PMID: 17406504
DOI: 10.1038/nprot.2006.484

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.

MeSH terms

Fluorescent Dyes / chemical synthesis*
Fluorescent Dyes / chemistry
Fluorescent Dyes / isolation & purification
Molecular Probe Techniques*
RNA / chemistry*

Substances

Fluorescent Dyes
RNA