Derivatization of controlled pore glass beads for solid phase oligonucleotide synthesis.
An improved and simplified procedure for the attachment of nucleosides onto long chain alkylamine controlled pore glass beads (LCAA-CPG) is presented. This procedure uses 1-(3-dimethylamino-propyl)-3-ethylcarbodiimide (DEC) to couple nucleoside 3'-succinates directly to the LCAA-CPG. The preparation of nucleoside 3'-succinate anhydrides, p-nitrophenyl, or pentachlorophenyl esters and the use of highly toxic dicyclohexylcarbodiimide (DCC) is no longer required. Procedures involving acidic activation of the LCAA-CPG before derivatization and a pre-synthesis capping are also described, which prevent the formation of oligonucleotides linked by 3'-phosphates to the LCAA-CPG. Evidence is presented indicating that this type of linkage is responsible for the apparently greater than 100% coupling yields observed for the first coupling cycle.
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Dept. of Medical Biochemistry, University of Calgary, Alberta, Canada.
Usman
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N
Ogilvie
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KK
D003124:Colorimetry; D004952:Esters / Q000138:chemical synthesis; D005898:Glass; D009705:Nucleosides / Q000138:chemical synthesis; D009841:Oligonucleotides / Q000138:chemical synthesis*; D011940:Receptor Aggregation; D013386:Succinates / Q000138:chemical synthesis
D004952:Esters; D009705:Nucleosides; D009841:Oligonucleotides; D013386:Succinates