Coupling: Use either dmf-dG (10-1029-xx) with standard monomers and Cap A or use UltraMILD monomers (dA: 10-1601-xx, dC: 10-1015-xx, dG: 10-1621-xx, dT: 10-1030-xx ) and UltraMILD Cap Mix A (40-4210-xx/ 40-4212-xx).
Deprotection: If standard monomers were used, deprotect in tert-butylamine/water 1:3 (v/v) for 6 hours at 60 °C. If UltraMILD monomers were used, deprotect in 0.05M Potassium Carbonate in Methanol, 4 hours at Room Temperature. See Technical Bulletin for details (http://www.glenresearch.com/Technical/TB_UltraMild_Deprotection.pdf). Note: A side reaction can occur that leads to a loss of the TAMRA label. Treatment of the support prior to cleavage/deprotection with a hindered base, such as 10% diethylamine in ACN, for 3-5 minutes will minimize the loss of label.
Storage: Freezer storage, -10 to -30°C, dry. Light sensitive material.
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%