In solid-phase synthesis, the oligonucleotides to be assembled are covalently bound to a solid support material by their 3'-terminal hydroxy groups and the solid support is packed in a column whose size depends on the scale of synthesis. The overwhelming majority of oligonucleotides are synthesized on small scale in columns ranging from 40 nmol to 10 mmol. The main functions of a synthesis column include maintaining a well-packed bed of solid support and ensuring the uniform distribution of phosphamides and reagents to all beads in the packed bed.
Once the desired full-length oligonucleotide has been successfully synthesized, the oligo chain is cleaved from the solid support with concentrated ammonium hydroxide, and the base-protecting groups are removed by heating in the ammonium hydroxide solution. Deprotected oligonucleotides can be purified and isolated by a variety of methods, such as precipitation, sizing columns, reversed-phase cartridges, denaturing urea polyacrylamide gel electrophoresis (PAGE), and liquid chromatography. The method chosen depends on molecular weight of oligonucleotides, purity required, and time considerations.
Amerigo Scientific offers accessories for synthesis of oligonucleotides, cartridges and barrels for the purification and desalting of oligonucleotides, nucleic acid purification systems, deprotection reagents and more.
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