Scientific Publications by FDA Staff
Curr Protoc Nucleic Acid Chem 2008 Dec;Chapter 3:Unit 3.17
Release of DNA oligonucleotides and their conjugates from controlled-pore glass under thermolytic conditions.
Grajkowski A, Cieslak J, Norris S, Freedberg DI, Kauffman JS, Duff RJ, Beaucage SL
The sequential functionalization of long-chain alkylamine controlled-pore glass (CPG) with a 3-hydroxypropyl-(2-cyanoethyl)thiophosphoryl linker and a dinucleoside phosphorotetrazolide leads to a uniquely engineered support for solid-phase synthesis. Unlike conventional succinylated-CPG supports, this support is designed to allow oligonucleotide deprotection and elimination of deprotection side-products to proceed without release of the oligonucleotide. When needed, the DNA oligonucleotide can be thermolytically released in 2 hr under essentially neutral conditions. The modified CPG support has been successfully employed in the synthesis of both native and fully phosphorothioated DNA 20-mers. On the basis of reversed-phase HPLC and electrophoretic analyses, the purity of the released oligonucleotides is comparable to that of identical oligonucleotides synthesized from succinylated-CPG supports, in terms of both shorter-than-full-length oligonucleotide contaminants and overall yields. The detailed preparation of DNA oligonucleotides conjugated with exemplary reporter or functional groups, either at the 3'-terminus or at both 3'- and 5'-termini, is also described.
|Category: Journal Article|
|PubMed ID: #19085983|
|Includes FDA Authors from Scientific Area(s): Drugs, Biologics|
|Entry Created: 2011-10-04||Entry Last Modified: 2012-08-29|