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Publisher Summary Microsphere-based platforms offer a high level of multiplexing capability and provide an “open” system in which reactions can be modified easily by combining different sets of reagent-bearing microspheres. With the completion of a representative human genomic sequence, emphasis is increasingly shifting toward rapid, cost-effective, high-throughput analysis of individual sequence variation. Although a portion of this variation occurs as nucleotide insertions and deletions, substitutions in the form of single nucleotide polymorphisms (SNPs) are the most common. Occasionally, a capture probe will consistently produce reaction products that display low signal strength or high backgrounds, resulting in poor signal-to-noise ratios. It is sometimes possible to correct this problem by redesigning the capture probe from the opposite genomic strand. Effective removal of unincorporated biotinylated dideoxynucleotides during the wash step can be critical to maximize signal-to-noise ratios. Microspheres can be photobleached by ambient laboratory light, causing them to lose their spectral identities. Because this is a cumulative process, long-term microsphere stocks, such as bulk carboxylated microspheres and oligonucleotide-coupled stocks, should be rigorously protected by wrapping in aluminum foil. |
