Some of your applications may require oligos of the highest quality. Cristallography studies, tri-dimensional construction, difficult cloning or gene synthesis are among these demanding applications.
The quality of the synthesis, the deprotection efficiency and the purification level are the most critical points to consider in order to reach the highest purity grade.
Eurogentec has developed an ultrahigh quality process to provide the best oligos from 8 to 140 bases long. This process relies on a proprietary synthesis and purification process combining a synthesis on polystyrene support, special amidites, optimized deprotection and double purification. This latter step is adapted based on the length and, if necessary, on the structure of the oligonucleotide.
- Optimized synthesis: the polystyrene 3’-end support has been selected for its high retention capacity and also its resistance to high amidites excess and long coupling times. This support is as efficient for long oligos as for short ones. Also the selected amidites are ultrapure increasing the coupling efficiency, at the same time the acetonitrile solvent is selected for its purity in order to optimize all reactions and washing steps.
- Improved deprotection: in order to provide a fully accessible oligo, without any protecting groups inactivating the final oligo product, we are applying a long, soft treatment allowing the optimal deprotection without damaging the sequence itself.
- Double purification: The purification will be selected mainly depending on the length of the oligo.
For an oligo between 8 and 39 bases long, a double HPLC will, in most of the cases, provide the best quality. This purification leads to 95 % purity.
For oligos between 40 and 140 bases long, a specific two-step purification process will be applied. First, a Reverse-Phase cartridge purification will eliminate most of the aborted sequences. A second “long PAGE” purification will allow very precise separation of the full-length oligo from the remaining n-x by-products. After this purification, a final gel filtration is performed to remove all the residual purification reagents. If required, a third round of purification will be performed in order to reach 95 % purity. - Quality Control : all UltraPureGold oligos are controlled in two stages. A first QC is performed before the purification in order to ensure that the synthesis was efficient enough to result in the 95 % purity expected for the final product. After the purification process a second quality control will check the quality of the final product.
This process has been co-developed with some customers and validated for many demanding applications. These purification steps are demanding on the final yield, therefore we cannot guarantee large final amounts of oligo products. But if your application requires a very high quality, this is the best process available on the market.
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