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Oligo Synthesis

Oligo Synthesis : Supports, CPGs

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Universal Support III

Universal Support III

Glen Research

Description

Universal Support III

Structure

 

Catalog Number: 20-5010-xx

(20-5010-xx, 20-5110-xx, 20-5210-xx)

Description: Universal Support III

1-Dimethoxytrityloxy-2-O-dichloroacetyl-propyl-3-N-succinyl-CPG
     

Diluent: Not Applicable
Coupling: This support should be used in a manner identical to normal protected nucleoside support since it contains the DMT group.
Deprotection: See Technical Bulletin for details (Technical Bulletin). Note: This product is not compatible with cyanoethyl protecting group removal protocols using hindered bases such as DEA or DBU. Removal of the cyanoethyl groups prior to cleavage will prevent the release of the oligonucleotide from the support.
Storage: Refrigerated storage, maximum of 2-8°C, dry
Stability in Solution: Not Applicable
Please Note: This product is covered by US Patent No.: 6,770,754.

Universal Support III

The key step in the use of any universal support in oligonucleotide synthesis is the dephosphorylation of the 3’-phosphate group to form the desired 3’-hydroxyl group. Azhayev1,2 has excelled in the investigation of neighboring group assistance in the dephosphorylation reaction. Amide groups may be considered to be weak N-H acids and can display basic properties in ammonium hydroxide or aqueous methylamine. (±)-3-Amino-1,2-propanediol was used to form a novel universal support. In our original US II support, a succinate linker attaches the 3-amino group to the support and the 2-OH is protected with a base-labile group to set up an amide assisted elimination in mildly basic conditions. In this way, the dephos­­phor­ylation reaction would eliminate the desired 3’-OH oligonucleotide into solution and the product of any ß-elimination competing side reaction would remain bound to the support. A further improvement has been achieved by using a carbamate group to connect the universal linker to the support, now called Universal Support III. The structures of the two supports are shown below right. Because the universal linker is unchanged and the succinate or carbamate groups remain attached to the support, we use the same catalog numbers for US II and III. Using Universal Support II or III, an oligo yield of > 80% can be achieved on CPG supports and > 95% on polymeric supports, with purity equivalent to the same oligo prepared normally.

Conditions for Cleavage and Deprotection are outlined in the table opposite. Universal Support II/III has been shown to generate oligonucleotides with the same efficacy in polymerase extension reactions as regular oligos. Despite the mild elimination reaction, oligonucleotides up to 75mer in length can be prepared routinely without loss of oligo during the synthesis cycles. This support is also used for the production of siRNA oligos.

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References

(1) A.V. Azhayev, Tetrahedron, 1999, 55, 787-800.

(2) A.V. Azhayev and M. Antopolsky, Tetrahedron, 2001, 57, 4977-4986.

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Notes

INTELLECTUAL PROPERTY

This product is covered by US Patent No.: 6,770,754 and European Patent No.: 1404695.

CLEAVAGE AND DEPROTECTION

Cleavage

For standard and UltraFast deprotection protocols, cleave the oligo from the support using 2M ammonia in methanol at room temperature for 30 minutes. (Only for oligonucleotides greater than 50 nucleotides in length, rinse the support with a further volume of water. Combine the two washes and evaporate to dryness.)

Deprotection

Standard

Add 1 volume of 30% ammonium hydroxide, seal and deprotect using the conditions appropriate for removal of the protecting groups on the nucleobases.

UltraFast

Add 1 volume of AMA (ammonium hydroxide/40% aqueous methylamine 1:1) seal and deprotect at 65°C for 10 minutes.

UltraMild Using Ammonium Hydroxide

Add 1 volume of ammonium hydroxide, seal and leave at room temperature for 8 hours.

UltraMild Cleavage and Deprotection
Using Potassium Carbonate in Methanol

Cleave the oligo from the support using 50 mM potassium carbonate in methanol at room temperature for 30 minutes. Seal and leave overnight at room temperature.

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