SYNTHESIS, DEPROTECTION, ANALYSIS AND PURIFICATION OF RNA AND RIBOZYMES

Improvements in the synthesis, deprotection and purification of oligor ibonucleotides are described, These advances allow for reduced synthes is and deprotection times, while improving product yield, Coupling tim es are reduced by half using 5-ethylthio-1H-tetrazole (S-ethyltetrazol e) as the activator, Base and 2'-O-t-butyldimethylsilyl deprotection w ith methylamine (MA) and anhydrous triethylamine/hydrogen fluoride in N-methylpyrrolidinone (TEA . HF/NMP), respectively, requires a fractio n of the time necessitated by current standard methods, In addition, t he ease of oligoribonucleotide purification and analysis have been sig nificantly enhanced using anion exchange chromatography. These new met hods improve the yield and quality of the oligoribonucleotides synthes ized, Hammerhead ribozymes synthesized utilizing the described methods exhibited no diminution in catalytic activity.