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.

Publisher's Statement

© The Royal Society of Chemistry 2014. Publisher’s version of record: https://doi.org/10.1039/c3ra46986g

Publication Title

RSC Advances

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