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An Efficient Method for the Isolation and Purification of Oligoribonucleotides |
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| Authors: | Brian Sproat Francesco Colonna Bashar Mullah Dean Tsou Alex Andrus Arnold Hampel |
| Institution: | 1. EMBL, Meyerhofstrasse 1 Heidelberg 69117 Germany Applied Biosystems Division Perkin Elmer Corporation 850 Lincoln Centre Drive , Foster City, CA, 94404, USA;2. CNR, Via P. Gobetti , 101, 40129, Bologna, Italy;3. Department of Biological Sciences , Northern Illinois University , DeKalb, IL, 60115, USA |
| Abstract: | Abstract Problems associated with the use of tetrabutylammonium fluoride like incomplete desilylation and removal of the tetrabutylammonium salts during large scale syntheses of oligoribonucleotides (RNA) have been eliminated by the use of triethylamine trihydrofluoride and precipitation of the RNA with 1-butanol. An efficient anion-exchange HPLC method has been developed for the purification of chemically synthesized RNA and the resulting product precipitated directly by the addition of 1-propanol. A new activator, 5-ethylthio-1H-tetrazole significantly enhances the synthesis quality and yield of oligoribonucleotides. RNA synthesized using these improvements has been shown to be biologically active by a comparative ribozyme-substrate assay. |
| Keywords: | NMR [5′‐13C]DNA C5′ resonance assignment C5′‐H1′ sequential assignment C5′‐H2″ sequential assignment 2D 1H‐13C HMQC‐NOESY |