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Methods and compositions for efficient nucleic acid sequencing

  • US 7,070,927 B2
  • Filed: 02/05/2002
  • Issued: 07/04/2006
  • Est. Priority Date: 09/27/1993
  • Status: Expired due to Fees
First Claim
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1. A method of determining a nucleotide sequence of a target nucleic acid, comprising the steps of:

  • (a) contacting a target nucleic acid with a set of immobilized oligonucleotide probe(s) and at least one labeled oligonucleotide probe from a set of labeled oligonucleotide probes under hybridization conditions effective to permit hybridization between;

    (i) complementary sequences of the target nucleic acid and the immobilized probes and (ii) complementary sequences of the target nucleic acid and the labeled probe(s);

    (b) covalently joining immobilized probe(s) and labeled probe(s) which are adjacently hybridized to the same target nucleic acid molecule;

    (c) detecting the labels of the labeled oligonucleotide probe(s) that are covalently joined to the immobilized probe(s);

    (d) identifying at least one nucleotide sequence in the target nucleic acid by steps comprising connecting the nucleotide sequences of the detected labeled oligonucleotide probe(s) with the nucleotide sequences of their respective joined immobilized oligonucleotide probe(s) (e) analyzing the sequences identified in step (d) to determine identified sequences that overlap and (f) reconstructing said sequence from overlapping oligonucleotide sequences identified in step (e).

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