Coupling: 15 minute coupling time recommended. Monomers that allow for UltraMILD deprotection are recommended. (Pac-A:10-3000-xx, Ac-C: 10-3015-xx, iPr-Pac-G: 10-3021-xx, U: 10-3030-xx ).
Deprotection: 2 M Ammonia in methanol, 24 hours at room temperature using UltraMILD phosphoramidites and UltraMILD Cap A is recommended. Alternatively, 60 hours at room temperature using standard phosphoramidites and Cap A will give acceptable results. To remove the 2’-TBDMS group, dry down the supernatant, treat with 1.5 mL of 1 M TBAF in THF for 18 hours at room temperature, quench with the addition of 1 mL of 1.5 M ammonium acetate and desalt (2x) using a Glen Gel-Pak™ 2.5 Desalting Column or equivalent.
Storage: Refrigerated storage, maximum of 2-8°C, dry
Stability in Solution: Similar to dA,C,G,T-CE Phosphoramidites
The table below shows pack size data and, for solutions, dilutions and approximate couplings based on normal priming procedures. Please link for more detailed usage information with the various synthesizers.