Recent highlights on photolithic oligonucleotide array in situ synthesis.
Identifieur interne : 002E86 ( Main/Exploration ); précédent : 002E85; suivant : 002E87Recent highlights on photolithic oligonucleotide array in situ synthesis.
Auteurs : Klaus-Peter Stengele [Allemagne] ; Jochen Bühler ; Sigrid Bühler ; Evgueni Kvassiouk ; Roland Green ; Tamara Prykota ; Wolfgang PfleidererSource :
- Nucleosides, nucleotides & nucleic acids [ 1525-7770 ] ; 2005.
Descripteurs français
- KwdFr :
- Analyse de profil d'expression de gènes (instrumentation), Biologie moléculaire (), Cinétique, Facteurs temps, Hybridation d'acides nucléiques, Modèles chimiques, Oligonucléotides (), Photochimie, Propanols (), Rayons ultraviolets, Sondes oligonucléotidiques, Séquençage par oligonucléotides en batterie (instrumentation), Techniques de chimie combinatoire.
- MESH :
- Analyse de profil d'expression de gènes, Biologie moléculaire, Cinétique, Facteurs temps, Hybridation d'acides nucléiques, Modèles chimiques, Oligonucléotides, Photochimie, Propanols, Rayons ultraviolets, Sondes oligonucléotidiques, Séquençage par oligonucléotides en batterie, Techniques de chimie combinatoire.
English descriptors
- KwdEn :
- Combinatorial Chemistry Techniques, Gene Expression Profiling (instrumentation), Kinetics, Models, Chemical, Molecular Biology (methods), Nucleic Acid Hybridization, Oligonucleotide Array Sequence Analysis (instrumentation), Oligonucleotide Probes, Oligonucleotides (chemistry), Photochemistry, Propanols (chemistry), Time Factors, Ultraviolet Rays.
- MESH :
- chemical , chemistry : Oligonucleotides, Propanols.
- chemical : Oligonucleotide Probes.
- instrumentation : Gene Expression Profiling, Oligonucleotide Array Sequence Analysis.
- methods : Molecular Biology.
- Combinatorial Chemistry Techniques, Kinetics, Models, Chemical, Nucleic Acid Hybridization, Photochemistry, Time Factors, Ultraviolet Rays.
Abstract
Light directed synthesis of high-density oligonucleotide microarrays is currently performed using either ortho-nitro-benzyl-type [MeNPOC] (Pease, A.C.; Solas, D.; Sullivan, E.J.; Cronin, T.M.; Holmes, C.P.; Fodor, S.P.A. Proc. Natl. Acad Sci U.SA. 1994, 91, 6333.) or ortho-nitrophenylethyl-type [NPPOC] (Hasan, A.; Stengele, K.P.; Giegrich, H.; Cornwell, P.; Isham, K.R.; Sachleben, R.A.; Pfleiderer, W.; Foote, R.S. Tetrahedron 1997, 53, 424Z) protecting groups as the 5'-O-carbonate ester of the phosphoramidite building block. The synthesis cycle uses a combinatorial approach attaching one specific base per cycle, thus as many as 100 cycles need to be run to make an array of 25-mers. Time needed for deprotection/activation of the growing oligo chain determines overall manufacturing time and consequently also cost. In this report we demonstrate the development of photoprotected posphoramidite monomers for light directed array synthesis with increasing sensitivity to the UV light used. If combined with maskless array synthesis, this technology allows for synthesis of arrays with >780,000 different 25-mer oligonucleotides in about one hour and allows for high flexibility in array design and reiterative redesign. The arrays synthesized show high quality and reproducibility in our standard hybridization based assay.
DOI: 10.1081/ncn-200059241
PubMed: 16248056
Affiliations:
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Prykota</name></author><author><name sortKey="Pfleiderer, Wolfgang" sort="Pfleiderer, Wolfgang" uniqKey="Pfleiderer W" first="Wolfgang" last="Pfleiderer">Wolfgang Pfleiderer</name></author></analytic><series><title level="j">Nucleosides, nucleotides & nucleic acids</title><idno type="ISSN">1525-7770</idno><imprint><date when="2005" type="published">2005</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Combinatorial Chemistry Techniques</term><term>Gene Expression Profiling (instrumentation)</term><term>Kinetics</term><term>Models, Chemical</term><term>Molecular Biology (methods)</term><term>Nucleic Acid Hybridization</term><term>Oligonucleotide Array Sequence Analysis (instrumentation)</term><term>Oligonucleotide Probes</term><term>Oligonucleotides (chemistry)</term><term>Photochemistry</term><term>Propanols (chemistry)</term><term>Time Factors</term><term>Ultraviolet Rays</term></keywords><keywords 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using either ortho-nitro-benzyl-type [MeNPOC] (Pease, A.C.; Solas, D.; Sullivan, E.J.; Cronin, T.M.; Holmes, C.P.; Fodor, S.P.A. Proc. Natl. Acad Sci U.SA. 1994, 91, 6333.) or ortho-nitrophenylethyl-type [NPPOC] (Hasan, A.; Stengele, K.P.; Giegrich, H.; Cornwell, P.; Isham, K.R.; Sachleben, R.A.; Pfleiderer, W.; Foote, R.S. Tetrahedron 1997, 53, 424Z) protecting groups as the 5'-O-carbonate ester of the phosphoramidite building block. The synthesis cycle uses a combinatorial approach attaching one specific base per cycle, thus as many as 100 cycles need to be run to make an array of 25-mers. Time needed for deprotection/activation of the growing oligo chain determines overall manufacturing time and consequently also cost. In this report we demonstrate the development of photoprotected posphoramidite monomers for light directed array synthesis with increasing sensitivity to the UV light used. If combined with maskless array synthesis, this technology allows for synthesis of arrays with >780,000 different 25-mer oligonucleotides in about one hour and allows for high flexibility in array design and reiterative redesign. The arrays synthesized show high quality and reproducibility in our standard hybridization based assay.</div></front></TEI><affiliations><list><country><li>Allemagne</li></country></list><tree><noCountry><name sortKey="Buhler, Jochen" sort="Buhler, Jochen" uniqKey="Buhler J" first="Jochen" last="Bühler">Jochen Bühler</name><name sortKey="Buhler, Sigrid" sort="Buhler, Sigrid" uniqKey="Buhler S" first="Sigrid" last="Bühler">Sigrid Bühler</name><name sortKey="Green, Roland" sort="Green, Roland" uniqKey="Green R" first="Roland" last="Green">Roland Green</name><name sortKey="Kvassiouk, Evgueni" sort="Kvassiouk, Evgueni" uniqKey="Kvassiouk E" first="Evgueni" last="Kvassiouk">Evgueni Kvassiouk</name><name sortKey="Pfleiderer, Wolfgang" sort="Pfleiderer, Wolfgang" uniqKey="Pfleiderer W" first="Wolfgang" last="Pfleiderer">Wolfgang Pfleiderer</name><name sortKey="Prykota, Tamara" sort="Prykota, Tamara" uniqKey="Prykota T" first="Tamara" last="Prykota">Tamara Prykota</name></noCountry><country name="Allemagne"><noRegion><name sortKey="Stengele, Klaus Peter" sort="Stengele, Klaus Peter" uniqKey="Stengele K" first="Klaus-Peter" last="Stengele">Klaus-Peter Stengele</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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