Solid-phase oligonucleotide synthesis and flow cytometric analysis with microspheres encoded with covalently attached fluorophores.
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.
Nanthakumar
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Axys Pharmaceuticals, 11099 North Torrey Pines Road, Suite 160, La Jolla, California 92037, USA.
Pon
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Mazumder
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Yu
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Watson
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D001896:Boron Compounds / Q000737:chemistry; D003432:Cross-Linking Reagents; D019075:Electrophoresis, Capillary; D005434:Flow Cytometry / Q000379:methods; D005456:Fluorescent Dyes / Q000737:chemistry; D008863:Microspheres; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000138:chemical synthesis*; D012680:Sensitivity and Specificity; D014867:Water / Q000494:pharmacology
C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D001896:Boron Compounds; D003432:Cross-Linking Reagents; D005456:Fluorescent Dyes; D009841:Oligonucleotides; D014867:Water