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Method for pair-wise sequencing a plurity of target polynucleotides

  • US 7,754,429 B2
  • Filed: 10/05/2007
  • Issued: 07/13/2010
  • Est. Priority Date: 10/06/2006
  • Status: Active Grant
First Claim
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1. A method for pairwise sequencing of first and second regions in each of a plurality of target polynucleotides, the method comprising:

  • (a) providing a solid support having immobilised thereon a plurality of first template polynucleotides linked to the solid support at their 5′

    ends, wherein a 5′

    -end immobilised primer is hybridised to the 3′

    end of each of the first template polynucleotides;

    (b) obtaining sequence data from a first region of the first template polynucleotides;

    (c) extending the 5′

    -end immobilised primers to copy the first template polynucleotides to generate a plurality of second immobilised template polynucleotides;

    (d) removing the plurality of first template polynucleotides from the solid support; and

    (e) obtaining sequence data for a second region of each of the second immobilised template polynucleotides, wherein determining the sequences of the first and second regions of the first and second template polynucleotides achieves pairwise sequencing of said first and second regions of said target polynucleotides.

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