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PubMed · 11257537

Polymer support for exonucleolytic sequencing.

M Hinz·S Gura·B Nitzan·S Margel·H Seliger
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Abstract

Different kinds of particles were investigated for their potential use as supports for exonucleolytic sequence analysis. Composite beads composed of an unreactive polystyrene "core" and a "shell" of functionalized silica nanoparticles were found to best fulfill the various prerequisites. The biotin/streptavidin system was used for attachment of DNA to composite beads of 6 microm diameter. Applying M13 ssDNA in extremely high dilution (approximately 1 molecule versus 100 beads) with internal fluorescent labels, only a small fraction of beads was found to be associated with fluorescent entities, which likely correspond to a very small number of bound DNA molecules per particle. For better selection and transfer of DNA-containing beads into microstructures for exonuclease degradation the loading experiments were repeated with composite beads of 2.3 microm diameter. In this case a covalent bond was formed between carboxylate-functionalized beads and amino-terminated oligonucleotides, which were detected through external labelling with fluorescent nanoparticles interacting with biotinylated segments of the complementary strand.

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BiotinDNA, Single-StrandedFluorescent DyesMicrospheresPolymersSequence Analysis, DNAStreptavidinJournal ArticleResearch Support, Non-U.S. Gov't

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M Hinz, S Gura, B Nitzan, S Margel, H Seliger. 2001-04-13. Polymer support for exonucleolytic sequencing.. https://doi.org/10.1016/s0168-1656(00)00419-3

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