Attachment of Nucleosides to Solid‐Phase Supports

Specific step‐by‐step instructions are given for coupling nucleosides to LCAA‐CPG supports (supports consisting of a long‐chain alkylamine linked to controlled‐pore glass). Protocols are given for a succinic acid linker and a hydroquinone‐O,O′‐diacetic acid linker. The former is the most widely used linker arm, and the starting materials are widely available. The latter offers greater compatibility with base‐sensitive sequence modifications and great synthetic throughput because it can be cleaved under milder and faster conditions. Additional guidelines are given for selecting a linker arm and coupling protocol. Almost any application requiring synthetic oligonucletodies can be satisfied using one of these linker arms.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/143722/1/cpnc0302.pd

Solid‐Phase Supports for Oligonucleotide Synthesis

This unit begins with a discussion of the advantages and disadvantages of oligonucleotide synthesis using solid supports. The physical and chemical properties of solid‐phase supports are discussed in terms of their suitability for oligonucleotide synthesis. In addition, the unit outlines the properties of linkers used for transient or permanent attachment of properly protected nucleosides to the derivatized support, as well as strategies for coupling nucleosides to linkers and conditions for the release of synthetic oligonucleotides from specific supports.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/143613/1/cpnc0301.pd

Strategies for Oligoribonucleotide Synthesis According to the Phosphoramidite Method

Advances in oligoribonucleotide synthesis have lagged behind those in oligodeoxyribonucleotide synthesis because of the difficulty in identifying orthogonal protecting groups for the 2′‐ and 5′‐hydroxyls. Adaptation of the phosphoramidite method for DNA synthesis to RNA synthesis has greatly improved our understanding of RNA. It allows site‐specific introduction of modified nucleosides to any position in an RNA molecule, as well as introduction of variations at multiple sites in the molecule. This overview discusses issues that are relevant to RNA synthesis by the phosphoramidite approach, including supports used, activation of the ribonucleoside phosphoramidites, and protection of the nucleobase, phosphate, and 2′‐ and 5′‐hydroxyls.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/143793/1/cpnc0305.pd