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Synthesis, deprotection, analysis and purification of RNA and ribozymes.
- Source :
-
Nucleic acids research [Nucleic Acids Res] 1995 Jul 25; Vol. 23 (14), pp. 2677-84. - Publication Year :
- 1995
-
Abstract
- 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.
- Subjects :
- Base Sequence
Chromatography, High Pressure Liquid
Chromatography, Ion Exchange
Ethylamines
Hydrofluoric Acid
Indicators and Reagents
Methods
Methylamines
Molecular Sequence Data
Pyrrolidinones
RNA analysis
RNA isolation & purification
RNA, Catalytic analysis
RNA, Catalytic isolation & purification
Tetrazoles
RNA chemical synthesis
RNA, Catalytic chemical synthesis
Subjects
Details
- Language :
- English
- ISSN :
- 0305-1048
- Volume :
- 23
- Issue :
- 14
- Database :
- MEDLINE
- Journal :
- Nucleic acids research
- Publication Type :
- Academic Journal
- Accession number :
- 7544462
- Full Text :
- https://doi.org/10.1093/nar/23.14.2677