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PubMed · 10725106

Solid-phase oligonucleotide synthesis and flow cytometric analysis with microspheres encoded with covalently attached fluorophores.

A Nanthakumar·R T Pon·A Mazumder·S Yu·A Watson
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Abstract

A novel combinatorial approach to synthesize oligonucleotides on fluorescently encoded microspheres based on flow sorting and segmental solid-phase synthesis is described. BODIPY dyes were covalently attached to polystyrene (8.8 microm, 55% DVB) microsphere particles to generate four fluorescently encoded sets. 20-mer oligonucleotide sequences can be synthesized on these microspheres with yields comparable to conventional CPG supports (80% overall yield, average stepwise yield = 99%). The concept of segmental solid-phase synthesis by flow sorting was demonstrated by synthesizing unique 20-mer oligonucleotide sequences on each of four fluorescently encoded microsphere sets by including a flow sorting step (after first eight base additions) and flow cytometric detection of sequences synthesized on each microsphere set by hybridization with fluorescently labeled complementary sequence.

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Boron CompoundsCross-Linking ReagentsElectrophoresis, CapillaryFlow CytometryFluorescent DyesMicrospheresNucleic Acid HybridizationOligonucleotidesSensitivity and SpecificityWaterJournal ArticleResearch Support, Non-U.S. Gov't

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A Nanthakumar, R T Pon, A Mazumder, S Yu, A Watson. Solid-phase oligonucleotide synthesis and flow cytometric analysis with microspheres encoded with covalently attached fluorophores.. https://doi.org/10.1021/bc990145z

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