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Microarray synthesis and assembly of gene-length polynucleotides

  • US 8,058,004 B2
  • Filed: 06/22/2009
  • Issued: 11/15/2011
  • Est. Priority Date: 09/12/2002
  • Status: Active Grant
First Claim
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1. A process for creating a mixture of oligonucleotide sequences in solution comprising:

  • (a) synthesizing in situ or spotting a plurality of oligonucleotide sequences on a microarray device or bead device each having a solid or porous surface, wherein the plurality of oligonucleotide sequences are attached to the solid or porous surface and wherein each oligonucleotide sequence comprises a fragment of a target polynucleotide sequence and further comprises two flanking sequences, one at the 3′

    end and the other at the 5′

    end of each fragment, wherein each flanking sequence is from about 7 to about 50 bases and comprising a primer region and a sequence segment having a restriction enzyme cleavable site;

    (b) amplifying each oligonucleotide sequence using primers complementary to the primer regions of the flanking sequence to form a plurality of double stranded oligonucleotide sequences; and

    (c) cleaving the primer regions from the plurality of double stranded oligonucleotide sequences at the restriction enzyme cleavable sites, thereby to produce a plurality of fragments of the target polynucleotide sequence,wherein the plurality of fragments together comprise the target polynucleotide sequence.

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