Controlled-pore glass, solid phase

Fig. 13 Strategy for solid-phase synthesis of dshRNA. (i) DMTr protection of glycerol, (ii) Attachment of succinyl linker to glycerol. (((() Attachment of glycerol to long-chain alkylamine controlled-pore glass solid support through succinyl linker, (iv) Oligonucleotide synthesis on DNA/RNA synthesizer, (v) Cleavage from soUd support and deprotection. X nucleotide in duplex, Y nucleotide in loop [127]...

Haller, W. (1983). Application of Controlled Pore Glass in Solid Phase Biochemistry (W. Haller, ed.), pp. 535-597. Wiley, New York. 

Haller, W., Application of controlled pore glass in solid phase biochemistry, In W.H. Scouten, ed. Solid Phase Biochemistry New York Wiley, 1983 535-597. 

Even controlled-pore glass (CPG) could be successfully employed as solid support with (9-glycosyl trichloroacetimidates as glycosyl donors. Thus, limitations of solvents and reaction temperatures in the glycosylation step, as experienced with the Merrifield resin, are restricted to those observed in solution-phase synthesis. Therefore, regio- and stereocontrol of the glycosylation reactions should be available from well-established solution-phase methodologies. 

In solid-phase syntheses, oligonucleotides are usually synthesized in the 5 -direction from an immobilized 3 -terminus. The solid phase is generally silica or controlled pore glass (CPG), which has been... 

For the automated solid-phase synthesis of oligonucleotides, however, silica was found to be the support of choice [192-197]. Silica with large pore size (25-300 nm), so-called controlled pore glass (CPG), is generally used for this purpose. The main advantages of CPG, as compared with silica gel, are its more regular particle size and shape, and greater mechanical stability. 

Chemical DNA-synthesis Another way to gain DNA is the chemical solid phase synthesis on controlled pore glasses (CPG), i.e. the phosphortriester method. The synthesis starts with a single nucleoside that is protected... 

Heckel A, Mross E, Jung KH, Rademann J, Schmidt RR. Oligosaccharide synthesis on controlled-pore glass as solid phase material. Synlett 1998 2 171-173. 

Kool reported the synthesis of macrocyclic nucleotide-hybrid compounds (215a-h), putative inhibitors of the HCV polymerase, polymerase C NS5B. The compounds were prepared by solid phase synthesis on controlled pore glass. 

Pon RT, Usman N, Ogilvie KK. Derivatization of controlled pore glass beads for solid-phase oligonucleotide synthesis. Biotechniques 6 768-775. 

Damha MJ, Giannaris PA, Zabarylo S. An improved procedure for derivati-zation of controlled pore glass beads or solid-phase oligonucleotide synthesis. Nucleic Acids Res 18 3813-3821, 1990. 

Oligodeoxynudeotide synthesis today is virtually always carried out on a solid support. Initially, the polymeric solid supports were polystyrene cross-linked with divinylbenzene, but controlled-pore glass beads (CPG beads) with defined porosities are now used in preference. The 3 -hydroxyl of the first protected deoxynucleotide is precoupled to the solid support and then solid phase, multistage oligodeoxynudeotide synthesis may commence in the 3 5 direction, in direct analogy to SPPS (Figure 2.5). 

Figure 2.5 Solid Phase DNA Synthesis Cycle (Contd.). Most frequently used base protecting groups are shown Bz Af-6 benzoyl (adenine), W-4 benzoyl (cytosine) N-2 isobutyroyl (guanine). All are base sensitive. DNA chain is built up from 3 to 5 on controlled-pore glass (CPG) bead solid support. Post global deprotection and resin removal, the desired product oligo-/polydeoxynucleotide is then separated initially by precipitation by means of an agent such as ethanol and purified finally by reversed phase liquid chromatography, or ion exchange chromatography as appropriate (see later in Chapter 2).

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