DNA Nucleoside Controlled Pore Glass (CPG ) Media | Data Sheet - DS0010EN00 | |
Solid support for DNA synthesis |
| • Uniform pore size allows efficient synthesis of pure oligomers • High surface area generates high yield of oligomers • Solvent-compatible support eliminates swelling and shrinking during synthesis cycles |
Solid support for DNA synthesis
The ability for research laboratories to cost-effectively synthesize DNA and RNA molecules in-house has greatly improved over the course of the past two decades. Automated oligonucleotide synthesizers are now available in most laboratories. Readily accessible technology has allowed synthetic molecules to be pervasively used in sequencing and in polymerase chain reaction (PCR) amplification techniques. Phosphoramidite chemistry is generally used for the synthesis of the oligonucleotides, which is carried out by sequentially adding the desired nucleotide units to an immobilized oligonucleotide chain.
Millipore offers DNA Nucleoside CPG DNA-CPG) products for the solidphase synthesis of oligonucleotides using the phosphoramidite chemistry. These products consist of the DNA nucleosides, deoxyadenosine (dA), deoxycytidine (dC), deoxyguanosine dG) and deoxythymidine (dT), covalently attached to a CPG particle (a porous borosilicate material) ready for synthesis applications. The unique characteristics of Millipore’s CPG matrix provide users with several advantages, including high yields in the synthesis of pure oligomers. A laboratory’s increased capacity improves the development process of therapeutic drugs as well as diagnostic probes on DNA chips.
Products
Covalently attached through the 3’ position to a CPG matrix, nucleosides dA, dC, dG and dT are available for solid-phase DNA synthesis. These products are manufactured from Millipore’s LCA-CPG matrix, a derivatized CPG product. Millipore produces DNA-CPG media on particles with 500 Å and 1000 Å pore sizes. A CPG 500 Å matrix is recommended for synthesis of oligomers up to 50 bases in length and a CPG 1000 Å matrix for oligomers up to 150 bases.
Advantages of a CPG Matrix
CPG material is recognized and accepted as the solid phase of choice for DNA synthesis. Its high surface area and narrow pore size distribution provide high coupling efficiency and purity for oligonucleotide synthesis. Incompressible and inert, the rigid glass structure is compatible with aqueous and organic solutions. The matrices are unaffected by changes in a solvent system, eliminating risks of swelling. The pore size of the CPG particles can be selected according to the size of the oligonucleotides to be synthesized.
Oligonucleotide Synthesis
The following steps describe a general procedure for solid phase oligonucleotide synthesis using the phosphoramidite chemistry and DNA-CPG particles:
Detritylation
The dimethoxytrityl (DMT) group that protects the 5’-OH is removed by treatment with trichloroacetic acid. This frees the 5’-OH for reaction.
Addition
Tetrazole and the phosphoramidite are mixed as they enter the reaction chamber. These form a highly reactive species that rapidly reacts with the 5’-OH.
Capping
Acetic anhydride and dimethylaminopyridine are mixed and used to terminate the growing oligomer and cap any chains that did not react during the addition step.
Oxidation
The labile trivalent phosphorous linkage formed in the addition step is converted to the stable, pentavalent phosphorous linkage of biologically active DNA. The oxidation step completes one cycle of oligonucleotide synthesis. DNA synthesis can continue with the removal of the DMT group at the 5’-OH of the growing chain, starting another cycle of nucleotide addition.
Deprotection
After chain assembly is complete, the phosphate protecting groups are removed, the chains are uncoupled from the solid support (CPG matrix), and the base protecting groups are removed. Oligonucleotide cleavage
from CPG particles and deprotection are achieved by treatment with concentrated ammonium hydroxide.
Typical Loading for Oligonucleosides
| Product | Coupling Efficiency | Ligand Loading Range (μmoles/g) |
| CPG 1000 Å, dA, dC, dG or dT | ≥97.5% | 20 - 40 |
| CPG 500 Å, dA, dC, dG or dT | ≥97.5% | 30 - 50 |
Covalent Attachment of Oligonucleosides to LCA-CPG Matrices
Manufacturing Standards and Quality Assurance
Millipore recognizes the importance of providing regulatory support and meeting industry quality standards. All CPG products are manufactured in a dedicated facility certified to internationally recognized standards, BS EN ISO9001. The facility is subject to routine surveillance audit.
Ordering Information
Please contact Millipore for customized orders.
| Description | Pack Size (g) | Catalogue No. |
| dA Nucleoside-CPG 1000 Å, 74 - 125 μm | 20 | 183934420 |
| dA Nucleoside-CPG 1000 Å, 74 - 125 μm | 200 | 183934421 |
| dC Nucleoside-CPG 1000 Å, 74 - 125 μm | 20 | 183935420 |
| dC Nucleoside-CPG 1000 Å, 74 - 125 μm | 200 | 183935421 |
| dG Nucleoside-CPG 1000 Å, 74 - 125 μm | 20 | 183936420 |
| dG Nucleoside-CPG 1000 Å, 74 - 125 μm | 200 | 183936421 |
| dT Nucleoside-CPG 1000 Å, 74 - 125 μm | 20 | 183937420 |
| dT Nucleoside-CPG 1000 Å, 74 - 125 μm | 200 | 183937421 |
| dA Nucleoside-CPG 500 Å, 74 - 125 μm | 20 | 185934420 |
| dA Nucleoside-CPG 500 Å, 74 - 125 μm | 200 | 185934421 |
| dC Nucleoside-CPG 500 Å, 74 - 125 μm | 20 | 185935420 |
| dC Nucleoside-CPG 500 Å, 74 - 125 μm | 200 | 185935421 |
| dG Nucleoside-CPG 500 Å, 74 - 125 μm | 20 | 185936420 |
| dG Nucleoside-CPG 500 Å, 74 - 125 μm | 200 | 185936421 |
| dT Nucleoside-CPG 500 Å, 74 - 125 μm | 20 | 185937420 |
| dT Nucleoside-CPG 500 Å, 74 - 125 μm | 200 | 185937421 |