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Use of mass spectrometry, capillary electrophoresis, and gel electrophoresis for quality analysis of synthetic oligonucleotides: Perspectives from the ABRF Nucleic Acid Research Group

Journal article published in 2001 by M. E. Gunthorpe, J. W. Fox, K. M. Hager, K. S. Lilley, S. Scaringe, A. T. Yeung
This paper is available in a repository.
This paper is available in a repository.

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Preprint: policy unknown
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Postprint: policy unknown
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Published version: policy unknown

Abstract

In 1995, the Nucleic Acids Research Group (NARG) of the Association of Biomolecular Resource Facilities (ABRF) eval-uated the quality of common oligonucleotides synthesized at core facilities and determined that these facilities provided products of high quality. In a 1999 survey, it was determined that most academic DNA synthesis core facilities regularly provide "boutique oligos," or modified oligonucleotides; how-ever, the quality of these products has not been the focus of a specific study. The NARG therefore designed an internal study whereby a standard oligonucleotide (M13F) and mod-ified forms of the oligonucleotide, including: 5Biotin, 5 TET (tetrachlorinated 6-carboxyfluorescein), and an antisense oligonucleotide using phosphorothioate chemistry, were syn-thesized by NARG members' laboratories. The products were then analyzed by matrix-assisted laser desorption ion-ization–time of flight mass spectrometry (MALDI-TOF MS), electrospray ionization mass spectrometry (ESI-MS), capillary electrophoresis (CE), and urea-gel electrophoresis.The qual-ity of the synthesis of these oligonucleotides as assessed by these methods are reported in this work along with a dis-cussion about the utility of these various quality-assurance technologies. (J Biomol Tech 2001;12:16–24) KEY WORDS: capillary electrophoresis, gel electrophoresis, mass spectrometry, modified oligonucleotides, oligonu-cleotides, quality assurance.