Highly parallel oligonucleotide purification and functionalization using reversible chemistry

We have developed a cost-effective, highly parallel method for purification and functionalization of 5′-labeled oligonucleotides. The approach is based on 5′-hexa-His phase tag purification, followed by exchange of the hexa-His tag for a functional group using reversible reaction chemistry. These me...

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Bibliographic Details
Published in:	Nucleic acids research Vol. 40; no. 1; p. e4
Main Authors:	York, Kerri T., Smith, Ryan C., Yang, Rob, Melnyk, Peter C., Wiley, Melissa M., Turk, Casey M., Ronaghi, Mostafa, Gunderson, Kevin L., Steemers, Frank J.
Format:	Journal Article
Language:	English
Published:	England Oxford University Press 01-01-2012
Subjects:	Biotinylation Chemistry Techniques, Synthetic Chromatography, Liquid Histidine - chemistry Methods Online Oligonucleotides - chemical synthesis Oligonucleotides - chemistry Oligonucleotides - isolation & purification Oligopeptides - chemistry
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Summary:	We have developed a cost-effective, highly parallel method for purification and functionalization of 5′-labeled oligonucleotides. The approach is based on 5′-hexa-His phase tag purification, followed by exchange of the hexa-His tag for a functional group using reversible reaction chemistry. These methods are suitable for large-scale (micromole to millimole) production of oligonucleotides and are amenable to highly parallel processing of many oligonucleotides individually or in high complexity pools. Examples of the preparation of 5′-biotin, 95-mer, oligonucleotide pools of >40K complexity at micromole scale are shown. These pools are prepared in up to ~16% yield and 90-99% purity. Approaches for using this method in other applications are also discussed.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors.
ISSN:	0305-1048 1362-4962
DOI:	10.1093/nar/gkr910