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Microarray Synthesis and Assembly of Gene-Length Polynucleotides

  • US 20100124767A1
  • Filed: 06/22/2009
  • Published: 05/20/2010
  • Est. Priority Date: 09/12/2002
  • Status: Active Grant
First Claim
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1. A process for assembling a polynucleotide from a plurality of oligonucleotides comprising:

  • (a) synthesizing or spotting a plurality of oligonucleotide sequences on a microarray device or bead device having a solid or porous surface, wherein a first oligonucleotide is oligo 1 and a second oligonucleotide is oligo 2 and so on, wherein the plurality of oligonucleotide sequences are attached to the solid or porous surface, and wherein the first oligonucleotide sequence has an overlapping sequence region of from about 10 to about 50 bases that is the same or substantially the same as a region of a second oligonucleotide sequence, and wherein the second oligonucleotide sequence has an overlapping region with a third oligonucleotide sequence and so on;

    (b) forming complimentary oligo 1 by extending primer 1, wherein primer 1 is complimentary to oligo 1;

    (c) disassociating complimentary oligo 1 from oligo 1 and annealing complimentary oligo 1 to both oligo 1 and to the overlapping region of oligo 2, wherein the annealing of complimentary oligo 1 to oligo 2 serves as a primer for extension for forming complimentary oligo 1+2;

    (d) repeating the primer extension cycles of step (c) until a full-length polynucleotide is produced; and

    (e) amplifying the assembled complementary full length polynucleotide to produce a full length polynucleotide in desired quantities.

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