Multiplexed differential displacement for nucleic acid determinations
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United States of America Patent
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Issued Date -
Apr 18, 2002
app pub date -
Aug 13, 2001
filing date -
Jul 16, 1999
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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. The subject method finds application for nucleic acid sequencing, single nucleotide polymorphism determinations, identification of nucleic acid fragments, and the like.
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