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SYSTEMS AND METHODS FOR MULTIPLEX ANALYSIS OF PCR IN REAL TIME

  • US 20080305481A1
  • Filed: 12/13/2007
  • Published: 12/11/2008
  • Est. Priority Date: 12/13/2006
  • Status: Active Grant
First Claim
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1. A method of amplifying and detecting a plurality of nucleic acid targets in a sample comprising:

  • (a) combining in a chamber a sample comprising the plurality of nucleic acid targets, a plurality of primer pairs for priming amplification of the plurality of nucleic acid targets, a labeling agent, and a first plurality of probes complementary to the plurality of nucleic acid targets, wherein the probes are immobilized on a plurality of encoded magnetic beads such that the identity of each probe is known from the encoded magnetic bead on which it is immobilized;

    (b) performing an amplification cycle to form labeled amplification products for each of the plurality of nucleic acid targets amplified with the plurality of primer pairs;

    (c) hybridizing the labeled amplification products to the probes immobilized on the encoded magnetic beads;

    (d) applying a magnetic field to a surface of the chamber to draw the encoded magnetic beads and the labeled amplification products hybridized to the probes immobilized on the encoded magnetic beads to the surface of the chamber;

    (e) detecting a signal from the encoded magnetic beads and a signal from the labeled amplification products;

    (f) removing the magnetic field from the surface of the chamber prior to performing a further amplification cycle; and

    (g) repeating steps (b) through (f) at least once;

    wherein the plurality of nucleic acid targets in the sample are amplified and detected.

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