Controlled Pore Glass (CPG) serves as a fundamental solid support in the synthesis of oligonucleotides, short DNA or RNA molecules crucial for various laboratory applications such as PCR, DNA sequencing, gene editing, and molecular biology research.
The process typically unfolds as follows:
Solid Support: CPG beads possess porous structures and a high surface area, making them ideal for the attachment of nucleotide building blocks during synthesis. This porous nature facilitates easy access for reagents to interact with the surface.
Protection and Deprotection: Oligonucleotide synthesis commences with the attachment of the initial nucleotide to the CPG support. Each nucleotide is strategically protected to prevent undesired reactions. Before adding the subsequent nucleotide, the protecting group is selectively removed. This iterative process is repeated for each nucleotide within the sequence.
Coupling Reaction: A crucial coupling reaction is executed to bind each nucleotide to the growing chain. In this step, the protected nucleotide undergoes activation and reacts with the free hydroxyl group of the preceding nucleotide attached to the CPG support. This reaction forms a phosphodiester bond, extending the oligonucleotide chain.
Washing and Purification: Post each coupling step, surplus reagents and by-products are thoroughly washed away to ensure purity. This meticulous step is pivotal in averting contamination and eliminating unreacted nucleotides.
Cleavage and Deprotection: Upon the synthesis of the desired sequence, the oligonucleotide is cleaved from the solid support. Concurrently, protecting groups are removed from the nucleotides, exposing their reactive sites.
Purification: The crude oligonucleotide undergoes purification utilizing various techniques such as high-performance liquid chromatography (HPLC) or polyacrylamide gel electrophoresis (PAGE). These methods effectively separate the oligonucleotide from impurities and truncated sequences.
Characterization: The final product undergoes characterization employing methods like UV spectroscopy to verify its purity and ascertain its concentration.
CPG-based synthesis presents several advantages including high efficiency, scalability, and compatibility with automated synthesis platforms. Widely adopted in research laboratories and biotech industries for oligonucleotide production, at SBS Genetech, our Controlled Pore Glass (CPG) is manufactured utilizing advanced production technology. This ensures precise processing and optimized control, resulting in superior particle size, shape, pore size, pore volume, and specific surface area. Such optimization endeavors to maximize nucleic acid purity and yield. Our CPG product stands as the gold standard solid-phase carrier in specialized fields, garnering extensive recognition and application.