Chemical capping by phosphitylation during oligonucleotide synthesis
AssigneeApplied Biosystems, LLC
InventorsWilliam A. Andrus, J. William Efcavitch, Lincoln J. McBride
Life of this patent
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Abstract
A method is provided for capping failure sequences in oligonucleotide synthesis by phosphitylation. A phosphite monoester is reacted with the 5' or 3' hydroxyl of the failure sequence between successive condensation steps in a synthesis procedure to form a 5' or 3' phosphite diester with the failure sequence. The phosphite diester substituent is inert with respect to subsequent reaction steps in the synthesis of the desired oligonucleotide product.
Claims16
A method of capping failure sequences in solid phase oligonucleotide synthesis, the method comprising the step of condensing a capping agent with a hydroxyl of a failure sequence, the capping agent being defined by the formula:
- ##STR5## wherein R is straight-chain, branch, or cyclic lower alkyl containing from 1 to 6 carbon atoms, electron withdrawing substituted lower alkyl, lower alkyl-substituted or halo-substituted aryl, or a nitrogen-, oxygen-, or sulfur-containing heterocycle having from 5 to 8 carbon atoms.
A method of synthesizing an oligonucleotide of a predetermined sequence on a solid support, the method comprising the steps of:
- (a) deprotecting a 5'-protected oligonucleotide attached to the solid support to form a deprotected oligonucleotide;
- (b) reacting a 5'-protected nucleotide monomer with the deprotected oligonucleotide to form either a 5'-protected oligonucleotide or a failure sequence, the failure sequence having a 5' hydroxyl;
- (c) capping the failure sequence by reacting a capping agent with the 5' hydroxyl of the failure sequence, the capping agent being defined by the formula:
- ##STR8## wherein R is lower alkyl, electron-withdrawing substituted lower alkyl, or lower alkyl- or halo-substituted aryl;
- and
- (d) repeating steps (a)-(c) until the oligonucleotide of the predetermined sequence is obtained.
Dependencies are inferred from the claim text.