Spacer Modification

Applicable Products

Physical & Dilution Data

Dilution volumes (in ml) are for 0.1M solutions in dry acetonitrile (4050). Adjust accordingly for other concentrations. For µmol pack sizes, products should be diluted as 100µmol/ml to achieve 0.1M, regardless of molecular weight.

Dissolution

Prepare the amidite solutions 5-10min before use. It is recommended these are vortexed to ensure complete dissolution before placing on the synthesiser.

Coupling, Cleavage & Deprotection

For all spacer phosphoramidites, no changes are required from standard synthesiser protocols for either coupling. Likewise, standard oligonucleotide deprotection conditions can be applied when deprotecting an oligo containing these modifications.

The CPGs are used in a manner identical to standard protected nucleoside supports, however non-nucleosidic modification (using 2245, 2395 or 2459) requires an initial detritylation prior to use in synthesis. For these modifications it is advised to carry out an initial deblock step, but not an initial capping step. This minimises the formation of N-1 product at the 3’ end. Cleavage and deprotection is achieved using the protocol required by the nucleobases. The cleavage and deprotection can also be done in one step by placing the resin in the deprotection solution then using the conditions required for deprotecting the nucleobases.

Storage & Stability

The products are stored dry in a freezer at –10 to –30°C. Phosphoramidite stability in solution is 2-3 days.