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Linkers and co-coupling agents for optimization of oligonucleotide synthesis and purification on solid supports

  • US 8,053,187 B2
  • Filed: 10/04/2010
  • Issued: 11/08/2011
  • Est. Priority Date: 03/14/2001
  • Status: Active Grant
First Claim
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1. A method of providing complementary nucleic acid comprising:

  • a) providing oligonucleotides, wherein said oligonucleotides are generated according to the method of;

    A) providing;

    i) a substrate comprising an array of oligonucleotides attached to said substrate via anchor moieties attached to non-cleavable linkers attached to said substrate, wherein said anchor moieties comprise the structure —

    C(X)—

    C(Y), wherein X comprises —

    OPO2O—

    , Y is a nucleophile and wherein said structure is part of a ring moiety;

    ii) a deprotecting solution; and

    iii) and a wash solution;

    B) deprotecting said oligonucleotides with said deprotecting solution;

    C) washing said oligonucleotides with said wash solution; and

    D) cleaving said oligonucleotides at said anchor moieties to provide purified oligonucleotides;

    b) mixing said oligonucleotides with a polymerase and template strands having a complementary sequence to the oligonucleotide in a polymerization buffer; and

    c) recovering copies of complementary nucleic acid strands in the duplex with the template strand.

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