Fast deprotection of synthetic oligodeoxyribonucleotides using standard reagents under microwave irradiation.
Fast methods for the removal of permanent amide exo-cyclic protective groups widely used in phosphoramidite-method DNA synthesis are desirable for many genomics and proteomics applications. In this communication, we present a method for the deprotection of a range of N-acyl deoxyribonucleosides (T, dA Bz, dC Bz, dC Ac, dG ibu, dG PAC) and synthetic oligodeoxyribonucleotides, ranging in length from 5-mer to 50-mer. Oligodeoxyribonucleotides were synthesized using standard amide protecting groups (dA Bz, dC Bz, dG ibu) and phosphoramidite chemistry on cis-diol solid phase support. This deprotection method utilizes 29% aqueous ammonia solution at 170 degrees C for 5 minutes under monomode microwave irradiation at a 20-nmole reaction scale. Reaction products were analyzed by TLC, RP-HPLC, CE, ESI-MS, real-time PCR, agarose gel electrophoresis, and by DNA uracil glycosylase (UDG) and phosphodiesterase I (PDE) enzymatic digestions.
Culf
Adrian S
AS
Atlantic Cancer Research Institute, Moncton, New Brunswick, Canada. adrianc@canceratl.ca
Cuperlović-Culf
Miroslava
M
Laflamme
Mark
M
Tardiff
Bennett J
BJ
Ouellette
Rodney J
RJ
D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D004587:Electrophoresis, Agar Gel; D019075:Electrophoresis, Capillary; D007202:Indicators and Reagents / Q000737:chemistry*; D008872:Microwaves; D009838:Oligodeoxyribonucleotides / Q000737:chemistry*; D016133:Polymerase Chain Reaction; D021241:Spectrometry, Mass, Electrospray Ionization
D007202:Indicators and Reagents; D009838:Oligodeoxyribonucleotides