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Creation of functionalized microparticle libraries by oligonucleotide ligation or elongation

  • US 8,486,629 B2
  • Filed: 04/26/2006
  • Issued: 07/16/2013
  • Est. Priority Date: 06/01/2005
  • Status: Active Grant
First Claim
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1. A method of creating a carrier-displayed library of oligonucleotide probes from a carrier-displayed general purpose library of oligonucleotide probes in a single multiplex reaction by ligation of anchor probes attached to encoded carriers and analyte-specific capture probes, comprising:

  • a. providing a general purpose library of anchor oligonucleotide probes, wherein anchor probes having different sequences are attached to differently encoded carriers;

    b. providing an analyte specific library of capture oligonucleotide probes which include subsequences complementary to designated analyte sequences;

    c. providing a library of clamp probes, each member of the library having a unique sequence and a first subsequence complementary to a terminal subsequence of an anchor probe and a second subsequence complementary to a terminal subsequence of a capture probe such that for each of the clamp probes, their entire first subsequence participates in duplex formation with the complementary anchor oligonucleotide probes and their entire second subsequence participates in duplex formation with the capture oligonucleotide probes without any internal mismatches or terminal overhangs;

    d. providing conditions permitting formation of tertiary complexes of capture probes, anchor probes and clamp probes;

    e. incubating said tertiary complexes under conditions such that the capture and the anchor probe are ligated to each other to generate a single oligonucleotide, wherein no monomers are added during or after ligation, andf. washing to release the clamp probes from the tertiary complexes.

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