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Highly parallel oligonucleotide purification and functionalization using reversible chemistry.

ABSTRACT: 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.

SUBMITTER: York KT

PROVIDER: S-EPMC3245951 | biostudies-literature | 2012 Jan

REPOSITORIES: biostudies-literature

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Highly parallel oligonucleotide purification and functionalization using reversible chemistry.

York Kerri T KT Smith Ryan C RC Yang Rob R Melnyk Peter C PC Wiley Melissa M MM Turk Casey M CM Ronaghi Mostafa M Gunderson Kevin L KL Steemers Frank J FJ

Nucleic acids research 20111029 1

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 po ...[more]

PMID: 22039155

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