Synthetic oligodeoxynucleotide purification by capping failure sequences with a methacrylamide phosphoramidite followed by polymerization
Document Type
Article
Publication Date
1-20-2014
Abstract
Oligodeoxynucleotide (ODN) purification was achieved by capping failure sequences with a polymerizable methacrylamide phosphoramidite during automated synthesis, polymerizing the failure sequences into an acrylamide gel after cleavage and deprotection, and extraction of full-length sequences with water. The details regarding the technology including the capping efficiency of four polymerizable phosphoramidites, optimal capping time, diffusion speeds of ODN from gels with different cross-linking ratios to solution, and the efficiency of ODN extraction from gel were investigated. In addition, the technology was tested for purification of a long sequence and purification on larger scales. We also found that polymerization of failure sequences in a centrifuge tube in air did not affect purification results. Finally, we provided additional evidence that ODNs are stable under radical polymerization conditions by complete digestion of ODN followed by reversed-phase HPLC analysis of nucleosides.
Publication Title
RSC Advances
Recommended Citation
Pokharel, D. P.,
Yuan, Y.,
Fueangfung, S.,
&
Fang, S.
(2014).
Synthetic oligodeoxynucleotide purification by capping failure sequences with a methacrylamide phosphoramidite followed by polymerization.
RSC Advances,
17, 8746-8757.
http://doi.org/10.1039/c3ra46986g
Retrieved from: https://digitalcommons.mtu.edu/chemistry-fp/130
Publisher's Statement
© The Royal Society of Chemistry 2014. Publisher’s version of record: https://doi.org/10.1039/c3ra46986g