Sequencing method using magnifying tags

  • US 6,723,513 B2
  • Filed: 06/22/2001
  • Issued: 04/20/2004
  • Est. Priority Date: 12/23/1998
  • Status: Expired due to Fees
First Claim
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1. A method of magnifying a signal associated with one or more nucleotide bases of a nucleotide sequence in a target nucleic acid molecule comprising the steps of:

  • (A) treating said target nucleic acid molecule so that at least a region of said target nucleic acid molecule is converted into an adapter binding region which is a region suitable for binding an adapter molecule;

    wherein said adapter molecule comprises;

    (i) one or more magnifying tags, or (ii) a means for attaching one or more of said magnifying tags to said adapter binding region, wherein said magnifying tag(s) correspond to;

    (a) one or more nucleotide bases of the adapter binding region or (b) one or more nucleotide bases adjacent to the adapter binding region or (c) one or more nucleotide bases which overlap the adapter binding region;

    (B) binding said adapter molecule to at least a portion of said adapter binding region created in step (A) to form a nucleic acid molecule;

    adapter molecule complex;

    (C) optionally, ligating said target nucleic acid molecule to said adapter molecule such that at least said magnifying tag(s) remain associated with said target nucleic acid molecule and to form a ligated nucleic acid molecule;

    adapter molecule complex;

    (D) treating the resulting complex of step (B) or the resulting complex of step (C) so that at least another region of said target nucleic acid molecule is converted into a form suitable for binding another adapter molecule, wherein said another region comprises one or more nucleotide bases which are not associated with the magnifying tags of step (B); and

    thereafter (E) repeating steps (B) to (D), with the proviso that the adapter molecule in each cycle of steps (B) to (C) binds to a region adjacent to a region of said target nucleic acid molecule to which the adapter molecule of a previous cycle bound, or the adapter molecule in each cycle of steps (B) to (C) binds to a region which overlaps with a region of said target nucleic acid molecule to which the adapter molecule of a previous cycle bound, and wherein the magnifying tags of each cycle of steps (A) to (C) are ligated together, to thereby magnify said signal.

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