Synthesis, deprotection, analysis and purification of RNA and ribosomes
| Autor: | Francine Wincott, Anthony DiRenzo, Chris Shaffer, Susan Grimm, Danuta Tracz, Christopher Workman, David Sweedler, Carolyn Gonzalez, Stephen Scaringe, Nassim Usman |
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| Rok vydání: | 1995 |
| Předmět: |
Molecular Sequence Data
Tetrazoles RNA chemical synthesis Hydrofluoric Acid Methylamines RNA analysis Ethylamines Methods Genetics RNA Catalytic Base sequence Chromatography High Pressure Liquid chemistry.chemical_classification Base Sequence biology Ion exchange Ribozyme RNA Chromatography Ion Exchange Pyrrolidinones Enzyme chemistry Biochemistry biology.protein Indicators and Reagents |
| Zdroj: | Nucleic Acids Research. 23:2677-2684 |
| ISSN: | 1362-4962 0305-1048 |
| Popis: | Improvements in the synthesis, deprotection and purification of oligoribonucleotides are described. These advances allow for reduced synthesis and deprotection times, while improving product yield. Coupling times are reduced by half using 5-ethylthio-1H-tetrazole (S-ethyltetrazole) as the activator. Base and 2'-O-t-butyldimethylsilyl deprotection with methylamine (MA) and anhydrous triethylamine/hydrogen fluoride in N-methylpyrrolidinone (TEA.HF/NMP), respectively, requires a fraction of the time necessitated by current standard methods. In addition, the ease of oligoribonucleotide purification and analysis have been significantly enhanced using anion exchange chromatography. These new methods improve the yield and quality of the oligoribonucleotides synthesized. Hammerhead ribozymes synthesized utilizing the described methods exhibited no diminution in catalytic activity. |
| Databáze: | OpenAIRE |
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