Coupling: 3 minute coupling time recommended. Use 0.02M Iodine to avoid degradation of the cyanine dye.
Deprotection: If dmf-dG is used during synthesis, deprotect in Ammonium Hydroxide for 2 hours at 65 °C; with ibu-dG, deprotect for 24-36 hours at room temperature. Note Cyanine 3 is compatible with UltraFast deprotection (AMA 10 minutes at 65 °C) if Ac-dC was used.
QUESTION: Does AMA or methylamine cause any degradation to fluorescein or fluorescein-type dyes such as FAM or FITC?
RESPONSE:Response: While AMA (Ammonium hydroxide/40% Methylamine 1:1 v/v) is considered compatible with fluorescein, the use of methylamine when deprotecting a Fluorescein-labeled oligo does lead to a small amount of degradation, which is characterized by a the appearance of a late-eluting peak by RP HPLC that shows no visible fluorescein absorbance. With standard deprotection conditions (AMA 10 minutes at 65 C) the amount of degradation is approximately 5%
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.