Advances in the chemical synthesis and purification of RNA.
Significantly developments that improve the chemical synthesis and purification of oligoribonucleotides have been attained. Introduction of a 2'-O-acetyl function on the nucleoside bound to the polystyrene support enhanced the yield of the oligoribonucleotide after cleavage. Higher coupling efficiency was achieved by the activation of phosphoramidites with a 0.75M solution of 5-ethylthio-1 H-tetrazole. Removal of the 2'-O-t-butyldimethylsilyl groups was effected rapidly at elevated temperatures with triethylamine trihydrofluoride in DMF. Fully deprotected RNA was then directly desalted and precipitated by the addition of 1-butanol. Purified RNA was isolated by the addition of 1-propanol to the collected HPLC purified fractions thus avoiding laborious desalting and solvent evaporation processes. These advancements have been extended to synthesis of ribozymes and tRNA.
Vinayak
R
R
Applied Biosystems Division, Perkin Elmer Corporation, Foster City, CA 94404, USA.
Andrus
A
A
Mullah
B
B
Tsou
D
D
D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D007202:Indicators and Reagents; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009843:Oligoribonucleotides / Q000138:chemical synthesis / Q000737:chemistry / Q000302:isolation & purification; D012313:RNA / Q000138:chemical synthesis* / Q000737:chemistry / Q000302:isolation & purification*
D007202:Indicators and Reagents; D009843:Oligoribonucleotides; D012313:RNA