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Reversible Biotinylation of the 5′‐Terminus of Oligodeoxyribonucleotides and its Application in Affinity Purification
dc.contributor.authorFang, Shiyue
dc.contributor.authorBergstrom, Donald E.
dc.date.accessioned2018-05-15T20:16:13Z
dc.date.available2018-05-15T20:16:13Z
dc.date.issued2003-09
dc.identifier.citationFang, Shiyue; Bergstrom, Donald E. (2003). "Reversible Biotinylation of the 5′‐Terminus of Oligodeoxyribonucleotides and its Application in Affinity Purification." Current Protocols in Nucleic Acid Chemistry 14(1): 4.20.1-4.20.17.
dc.identifier.issn1934-9270
dc.identifier.issn1934-9289
dc.identifier.urihttps://hdl.handle.net/2027.42/143794
dc.description.abstractThe preparation of two reversible biotinylation phosphoramidites and their application in labeling and affinity purification of synthetic oligodeoxyribonucleotides will be described. In both cases, the biotin is linked to the 5′‐terminus of DNA through a diisopropyl silyl acetal functionality. This linkage is completely stable under certain postsynthetic cleavage/deprotection conditions, but can be readily broken by fluoride ions, releasing unmodified 5′‐OH and 5′‐phosphate DNA, respectively. To demonstrate the use of these reversible biotinylation methods, crude DNA was incubated with NeutrAvidin‐coated microspheres, full‐length biotinylated DNA was efficiently attached to the solid phase, and nonbiotinylated failure sequences and other impurities were readily removed by washing with buffer. Cleavage of the silyl acetal linkage afforded high‐quality, full‐length, unmodified 5′‐OH and 5′‐phosphate DNA, respectively, depending on which of the two phosphoramidites was used. It is anticipated that this method will find applications in areas that require efficient isolation of DNA from a complex mixture.
dc.publisherJohn Wiley & Sons
dc.subject.otherDNA
dc.subject.otherAvidin
dc.subject.otherSilyl Acetal
dc.subject.otherFluoride Cleavage
dc.subject.otherBiotinylation
dc.subject.otherPhosphoramidite
dc.subject.otherAffinity Purification
dc.titleReversible Biotinylation of the 5′‐Terminus of Oligodeoxyribonucleotides and its Application in Affinity Purification
dc.typeArticleen_US
dc.rights.robotsIndexNoFollow
dc.subject.hlbsecondlevelPublic Health
dc.subject.hlbsecondlevelBiological Chemistry
dc.subject.hlbsecondlevelChemical Engineering
dc.subject.hlbsecondlevelChemistry
dc.subject.hlbtoplevelHealth Sciences
dc.subject.hlbtoplevelScience
dc.subject.hlbtoplevelEngineering
dc.description.peerreviewedPeer Reviewed
dc.description.bitstreamurlhttps://deepblue.lib.umich.edu/bitstream/2027.42/143794/1/cpnc0420.pdf
dc.identifier.doi10.1002/0471142700.nc0420s14
dc.identifier.sourceCurrent Protocols in Nucleic Acid Chemistry
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dc.owningcollnameInterdisciplinary and Peer-Reviewed


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