Linker phosphoramidite reagents for the attachment of the first nucleoside to underivatized solid-phase supports.
New linker phosphoramidite reagents containing a cleavable 3'-ester linkage are used for attaching the first nucleoside to the surface of a solid- phase support. Inexpensive, underivatized amino supports, such as long chain alkylamine controlled-pore glass, can serve as universal supports. No modifications to phosphoramidite coupling conditions are required and, after synthesis, treatment with NH(4)OH releases the products with 3'-OH ends. No 3'-dephosphorylation is required. Phosphoramidite reagents containing a succinate and sulfonyl diethanol linkage between the nucleoside and phosphoramidite group are particularly advantageous and can be used to create both 3'-OH and 5'-phosphate ends on oligonucleotides. Reproducibility and quality of oligonucleotide synthesis is demonstrated for either column and 96-well plate formats on low-, medium- or high-loading CPG supports.
Pon
Richard T
RT
Department of Biochemistry and Molecular Biology, University of Calgary, 3550 Hospital Drive N.W., Calgary, Alberta, Canada T2N 4N1. rtpon@ucalgary.ca
Yu
Shuyuan
S
D000588:Amines / Q000737:chemistry; D001483:Base Sequence; D004952:Esters / Q000737:chemistry; D005898:Glass / Q000737:chemistry*; D009705:Nucleosides / Q000737:chemistry*; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis* / Q000737:chemistry*; D009943:Organophosphorus Compounds / Q000737:chemistry*; D010766:Phosphorylation; D015203:Reproducibility of Results
D000588:Amines; D004952:Esters; D009705:Nucleosides; D009838:Oligodeoxyribonucleotides; D009943:Organophosphorus Compounds; C434331:phosphoramidite