Attachment of Nucleosides and Other Linkers to Solid‐Phase Supports for Oligonucleotide Synthesis

Abstract

Specific step-by-step instructions for conversion of 5'-O-(4,4'-dimethoxytrityl)- and base-protected nucleosides and other mono-O-(4,4'-dimethoxytrityl)-protected diols to their hemisuccinate esters and their coupling to CPG (controlled-pore glass) supports bearing aminopropyl or long chain aminoalkyl groups are presented. Additional guidelines are provided for selecting a coupling protocol and performing in-process control.

Circular DNA by “Bis‐Click” Ligation: Template‐Independent Intramolecular Circularization of Oligonucleotides with Terminal Alkynyl Groups Utilizing Bifunctional Azides

“…The CPG support was filtered off and washed successively with pyridine (5 mL), methanol (10 mL), and dichloromethane (20 mL), and air‐dried at room temperature. The amount of CPG‐bound nucleoside residues was determined by detritylation of the CPG beads . A sample of CPG conjugate (4 mg) was treated with dichloroacetic acid (10 mL; 2.5 % in dry CH 2 Cl 2 ) for 10 min.…”

Synthesis of 2′-O-Methyl/2′-O-MOE-L-Nucleoside Derivatives and Their Applications: Preparation of G-Quadruplexes, Their Characterization, and Stability Studies

“…The l -nucleoside succinate derivatives 29 and 30 were obtained with an 93 and 91% yield, respectively. These compounds were used for the functionalization of long-chain aminoalkyl controlled pore glass (LCAA-CPG) supports in a conventional manner to yield the CPG solid-supports 31 and 32 with 20 and 23 μmol/g loading, respectively …”

“…Thus, the addition of tertbutylhydroperoxide to a mixture of compound 12 and NaSO 2 CF 3 in water provided 2′-O-methyl-5-trifluoromethyl- L-uridine (25,Scheme 5) in 29% yield after 26 h of reaction at rt. In a similar way, compound 13 was transformed into 2′-O-MOE-5-trifluoromethyl-L-uridine (26) in modest yield.…”

“…These compounds were used for the functionalization of long-chain aminoalkyl controlled pore glass (LCAA-CPG) supports in a conventional manner to yield the CPG solid-supports 31 and 32 with 20 and 23 μmol/g loading, respectively. 26 The synthesis of phosphoramidites derived from 2′substituted L-uridine was carried out starting from the corresponding 5′-O-DMT-protected analogues (Scheme 6). Phosphitylation of 27 and 28, with chloro-2-cyanoethoxy-N,Ndiisopropylaminophosphine gave the expected phosphoramidites 33 and 34 in 67 and 65% yield, respectively.…”

A Simple and Efficient Microfluidic System for Reverse Chemical Synthesis (5′-3′) of a Short-Chain Oligonucleotide Without Inert Atmosphere

“…The spectrophotometric assay of the resultant solution (containing trityl cations) was collected and measured to monitor the efficiency of the synthesis step. Stepwise coupling yields were estimated on the basis of the characteristic trityl absorbance assay in CH 2 Cl 2 (ε = 76000 LM −1 cm −1 ) (see supplementary information, 9) [25]. The residual deblocking reagent in the chamber was washed out through the outlet via a 2-3 min superdehydrated acetonitrile washing step performed at the same flow rate.…”