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REVERSIBLE DI-NUCLEOTIDE TERMINATOR SEQUENCING

  • US 20090263797A1
  • Filed: 11/20/2008
  • Published: 10/22/2009
  • Est. Priority Date: 11/20/2007
  • Status: Active Grant
First Claim
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1. A method for determining a sequence of nucleotides in a template, the method comprising:

  • (a) hybridizing a primer to a template;

    (b) extending the primer in the presence of a reversible di-nucleotide terminator, wherein the reversible di-nucleotide terminator comprises a first nucleotide and a second nucleotide;

    (c) identifying a family of the reversible di-nucleotide terminator;

    (d) generating an extendable terminus on the incorporated reversible di-nucleotide terminator;

    (e) repeating steps (b), (c), and (d) a suitable number of times to form an extension product hybridized to the template;

    (f) removing the extension product;

    (g) hybridizing a shifted-primer to the template;

    (h) extending the primer in the presence of a reversible di-nucleotide terminator, wherein the reversible di-nucleotide terminator comprises a first nucleotide and a second nucleotide;

    (i) identifying a family of the reversible di-nucleotide terminator;

    j) generating an extendable terminus on the incorporated reversible di-nucleotide terminator;

    (k) repeating steps (h), (i), and (j) a suitable number of times; and

    ,(l) sequencing the template.

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