Method for loading solid supports for nucleic acid synthesis
First Claim
1. A method of loading a functionalized solid support for oligonucleotide synthesis comprising simultaneously contacting in a solvent the functionalized solid support with diisopropylcarbodiimide, a nucleoside having a 3'"'"' linker group attached, and an acid catalyst.
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Abstract
New methods of loading solid supports for oligonucleotide synthesis are presented. The new methods comprise coupling a succinylated nucleoside to a solid support using diisopropylcarbodiimide (DIC) as an activator and N-hydroxybenzotriazole (HOBT) as an acid catalyst. DIC is substantially cheaper than the currently used activating agent, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (DEC). Furthermore, in the preferred embodiment, when it is used in combination with HOBT, coupling is more efficient, requiring less nucleoside to achieve the same loading densities as in the prior art. The loading process is faster than prior art methods and the overall cost savings of about 43% are realized.
39 Citations
27 Claims
- 1. A method of loading a functionalized solid support for oligonucleotide synthesis comprising simultaneously contacting in a solvent the functionalized solid support with diisopropylcarbodiimide, a nucleoside having a 3'"'"' linker group attached, and an acid catalyst.
- 14. A method of loading a functionalized solid support for oligonucleotide synthesis comprising simultaneously contacting in a solvent a functionalized solid support having a linker moiety with diisopropylcarbodiimide, a nucleoside having a free 3'"'"' hydroxy group, and an acid catalyst.
Specification