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Issue 17, 2014
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Synthetic oligodeoxynucleotide purification by capping failure sequences with a methacrylamide phosphoramidite followed by polymerization

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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.

Graphical abstract: Synthetic oligodeoxynucleotide purification by capping failure sequences with a methacrylamide phosphoramidite followed by polymerization

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Publication details

The article was received on 24 Nov 2013, accepted on 17 Jan 2014 and first published on 20 Jan 2014


Article type: Paper
DOI: 10.1039/C3RA46986G
Author version available: Download Author version (PDF)
Citation: RSC Adv., 2014,4, 8746-8757
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    Synthetic oligodeoxynucleotide purification by capping failure sequences with a methacrylamide phosphoramidite followed by polymerization

    D. Pokharel, Y. Yuan, S. Fueangfung and S. Fang, RSC Adv., 2014, 4, 8746
    DOI: 10.1039/C3RA46986G

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