Protocol abstract
Nature Protocols 2, - 1052 - 1058 (2007)
Published online: 26 April 2007 | doi:10.1038/nprot.2007.154
Subject Categories: Isolation, purification and separation | Chemical modification | Nucleic acid based molecular biology | Synthetic chemistry | Spectroscopy and structural analysis
Synthesis of oligo-RNAs with photocaged adenosine 2'-hydroxyls
Steven G Chaulk1 & Andrew M MacMillan1
Abstract
This protocol describes a general method for the preparation of RNAs in which the reactivity or hydrogen-bonding properties of the molecule are modified in a photoreversible fashion by use of a caging strategy. A single caged adenosine, modified at the 2' position as a nitro-benzyl ether, can be incorporated into short RNAs by chemical synthesis or into long RNAs by a combination of chemical and enzymatic synthesis. The modified RNAs can be uncaged by photolysis under a variety of conditions including the use of a laser or xenon lamp, and the course of this uncaging reaction may be readily followed by HPLC or thin-layer chromatography.
- Department of Biochemistry, University of Alberta Edmonton, Alberta, Canada T6G 2H7.
Correspondence to: Andrew M MacMillan1 e-mail: andrew.macmillan@ualberta.ca
nature-products
A-Z product listing
- 2-Cyanoethyl-N,N-diisopropylaminochlorophosphite(Aldrich)
- 2-Nitrobenzylbromide(Aldrich)
- 2'-FPMP RNA phosphoramidites(Rasayan Inc.)
- 4,4'-Dimethoxytrityl chloride(Aldrich)
- 500 Å FPMP-controlled pore glass resin(Rasayan Inc.)
- [
-32P]ATP(NEN)
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