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Optimized primer library for gene sequencing and method of using same

  • US 6,083,695 A
  • Filed: 05/21/1997
  • Issued: 07/04/2000
  • Est. Priority Date: 04/15/1996
  • Status: Expired due to Fees
First Claim
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1. A method for sequencing an unknown DNA molecule comprising the steps of:

  • selecting a library primer from an octamer oligonucleotide library consisting of sequence ID numbers 5, 32, 36, 47, 54, 73, 79, 103, 116, 121, 125, 204, 248, 274, 283, 351, 354, 358, 371, 379, 380, 384, 387, 397, 401, 402, 403, 439, 440, 445, 450, 454, 457, 460, 496, 499, 500, 501, 517 519, 523, 528, 529, 544, 545, 552, 553, and 554 and the corresponding complementary sequences, wherein the library primer is complementary to a known sequence adjacent to the unknown sequence or is complementary to a sequence in a known extension product;

    adding the library primer to a single stranded version of the unknown DNA molecule;

    annealing the library primer to the single stranded unknown DNA molecule under conditions where the library primer anneals to an exactly matched complementary sequence in the unknown DNA molecule;

    synthesizing an extension product from the 3'"'"' end of the library primer said extension product complementary to the single stranded unknown DNA molecule;

    analyzing the extension product to determine the sequence; and

    repeating the selecting, adding, annealing, synthesizing and analyzing steps until the sequence of the unknown DNA molecule is determined.

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