Read below to understand all of the steps in oligonucleotide synthesis.
The process of oligonucleotide synthesis may seem daunting and complex but the procedure is somewhat basic. Because of the ability and versatility of the oligonucleotide, the steps that are taken are handled by the modified oligo itself. Here is a breakdown of the steps that occur during the oligo synthesis process.
The first step in the oligonucleotide synthesis process is adding a base that to ensure that the sites become unblocked for further use. The initial base which is attached to the solid support is inactive because of all the sites becoming blocked. By adding a dichloroacetic or a trichloroacetic acid, the hydroxyl group will be the only reactive group on the monomer.
The second step would be to activate the monomer and to adjoin two reactive bases together. The addition of tetrazole to the base protects the phosphorus linkage therefore activating the column.
The next step is more of a fail-safe procedure as there should not be an oligonucleotide created with a deletion. Active groups are capped with protective groups to prevent strands from growing again and causing there to be excessive binding.
The last step would be for the unstable phosphite linkage to be oxidized to form a stable base column. The steps are then all repeated until all of the bases have been added to the oligonucleotide. These steps work in congruence with each other to ensure that the rna polymers and dna polymers associated with the oligonucleotide bind in an appropriate fashion.