UNIT 11.11 Site-Specific Fluorescent Labeling of Large RNAs with Pyrene
Published Online: 1 DEC 2004
Copyright © 2004 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Nucleic Acid Chemistry
How to Cite
Smalley, M. K. and Silverman, S. K. 2004. Site-Specific Fluorescent Labeling of Large RNAs with Pyrene. Current Protocols in Nucleic Acid Chemistry. 19:11.11:11.11.1–11.11.12.
- Published Online: 1 DEC 2004
- Published Print: NOV 2004
Pyrene is a useful chromophore for monitoring the tertiary structure and folding of large RNAs. This unit describes the general preparation of a large RNA (>80 nucleotides in length) that has been site-specifically modified with pyrene at the 2′-position of an individual internal nucleotide. A protocol is provided for derivatizing a 2′-amino-RNA oligonucleotide with a suitably activated pyrene reagent. This pyrene-labeled oligonucleotide is then assembled with other RNA(s) either by covalent ligation or by noncovalent hybridization to form a full-length structured RNA, which may then be studied by equilibrium and stopped-flow fluorescence spectroscopy.
- RNA folding;