Solid phase oligonucleotide synthesis

The phosphoramidite of 8-Ph-dG has been prepared and used to site-specifically incorporate it into oligonucleotides using solid-phase DNA synthesis. In the 12-mer 5 -d(GCGCCXGCGGTG), where X = 8-Ph-dG, the adduct behaved very similar to 8-Me-dG with of the duplex being 5.6°C lower than the normal dG dC base pair, while the of 8-Ph-dG dG was 2.1°C higher than that of the mismatched dG dC base pair. This data suggested that, similar to 8-Me-dG, 8-Ph-dG exists primarily in the anti conformation when paired with dC, while the syn conformation may pair with dGMP. ... 

Sahl993 Saha, A.K., Sardaro, M., Waychunas, C., Delecki, D., Kruse, L.I., Kutny, R., Cavanaugh, R, Yawman, A. and Upson, D.A., Diisopropylsilyl-linked Oligonucleotide Analogs Solid-Phase Synthesis and Physicochemical Properties, J. Org. Chem., 58(1993) 7827-7831. 

Morvan, F., Rayner, B., and Imbach, J. L (1990) Modified oligonucleotides-IV. Solid phase synthesis and preliminary evaluation of phosphorothioate RNA as potential antisense agents Tetrahedron Lett 31,7149-7152. 

In order to achieve good-quality synthesis on a large scale, proper setting of the instrument is an integral part of the process. Here, large-scale synthesis of oligonucleotides on solid-phase has been discussed using a model 8800 DNA synthesizer. The instrument 8800 DNA synthesizer consists of different components ... 

F. Morvan, B. Rayner and J.L. Imbach, Modified oligonucleotides IV Solid phase synthesis... 

The protected nucleoside-3-phosphoramidite monomer units such as 671 are used in the solid-phase oligonucleotide synthesis. In the 60mer synthesis, 104 allylic protective groups are removed in almost 100% overall yield by the single Pd-catalyze reaction with formic acid and BuNH2[432], N,(9-protection of uridine derivatives was carried out under pha.se-transfer conditions[433]. 

Synthetic organic polymers, which are used as polymeric supports for chromatography, as catalysts, as solid-phase supports for peptide and oligonucleotide synthesis, and for diagnosis, are based mainly on polystyrene, polystyrene-divinylbenzene, polyacrylamide, polymethacrylates, and polyvinyl alcohols. A conventional suspension of polymerization is usually used to produce these organic polymeric supports, especially in large-scale industrial production. 

One widely used method involving protected compounds is solid-phase synthesis (polymer-supponed reagents). This method has the advantage of requiring only a simple workup by filtration such as in automated syntheses, especially of polypeptides, oligonucleotides, and oligosaccharides. 

Solid-phase chemistry is an efficient synthetic tool that, compared with solution-phase chemistry, simplifies the work-up of the reaction, allows the process to be driven to completion by using excess of reagents, and can be automatized [2a]. In recent years, many studies have been devoted to developing both surface-mediated and resin-supported synthesis. Today the solid-phase approach is not limited to peptides and oligonucleotides but is also used to synthesize molecules of lower molecular weight. 

Synthetic oligonucleotides are very important tools in the study and manipulation of DNA, including such techniques as site-directed mutagenesis and DNA amplification by the polymerase chain reaction. The techniques for chemical synthesis of oligonucleotides are highly developed. Very efficient automated methodologies based on solid phase synthesis are used extensively in fields that depend on the availability of defined DNA sequences.52... 

Silica or porous glass is usually used as the solid phase in oligonucleotide synthesis. The support is functionalized through an amino group attached to the silica surface. There is a secondary linkage through a succinate ester to the terminal 3 -OH group. 

Over the past decade the techniques of combinatorial synthesis have received much attention. Solid phase synthesis of polypeptides and oligonucleotides are especially adaptable to combinatorial synthesis, but the method is not limited to these fields. The goal of combinatorial synthesis is to prepare a large number of related... 

---