Abstract: Multiplexed determinations of large numbers of events are achieved in an accurate and simple manner by using a multitude of primer reagents in combination with different capture reagents that serve for sequestering all the reagents at a single site, followed by independent release of subsets of the primer reagents using differential release conditions. Also included as part of the primer reagents may be identifiers, which serve to identify a particular characteristic. The method is illustrated using primers with sequences for initiation of chain extension that are joined to or serve as a capture sequence, and where the extended primer has an identifier. After extending the primer, the extended primers are sequestered via the capture sequence onto a sequestering agent, sequentially released and the released extended primers analyzed to provide multiplexed determinations.

Abstract: Multiplexed determinations of large numbers of events are achieved in an accurate and simple manner by using a multitude of primer reagents in combination with different capture reagents that serve for sequestering all the reagents at a single site, followed by independent release of subsets of the primer reagents using differential release conditions. Also included as part of the primer reagents may be identifiers, which serve to identify a particular characteristic. The method is illustrated using primers with sequences for initiation of chain extension that are joined to or serve as a capture sequence, and where the extended primer has an identifier. After extending the primer, the extended primers are sequestered via the capture sequence onto a sequestering agent, sequentially released and the released extended primers analyzed to provide multiplexed determinations.

Abstract: Reagents capable of forming branched oligomers with monomeric units are disclosed, together with oligomers incorporating such reagents, kits containing such reagents and methods for use of such reagents in forming oligomers with monomeric units. The present reagents can advantageously be used to introduce multiple labels or reporter molecules onto oligomers such as oligonucleotides and oligopeptides. In particular, non-nucleosidic phosphoramidites based on 1,3,5-tris(2-hydroxyethyl)cyanuric acid are disclosed. Multiply-labeled, branched DNA oligomer probes constructed using these phosphoramidite reagents showed increased signal intensity relative to singly-labeled oligomer probes.