COMPOSITIONS AND METHODS FOR NUCLEIC ACID SEQUENCING

  • US 20130309678A1
  • Filed: 04/19/2013
  • Published: 11/21/2013
  • Est. Priority Date: 03/28/2008
  • Status: Active Grant
First Claim
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1. A method of determining a nucleotide sequence of a double stranded nucleic acid segment, comprising:

  • providing a nucleic acid molecule comprising a double-stranded portion, the double-stranded portion comprising a forward strand and a reverse strand of the double-stranded nucleic acid segment, wherein the forward strand and the reverse strand are complementary to each other;

    linking a 3 end of the forward strand to a 5′

    end of the reverse strand, thereby forming a sequencing template, wherein said 3′

    end of the forward strand and said 5′

    end of the reverse strand are linked by a linking oligonucleotide;

    performing single-molecule sequencing of the sequencing template, thereby obtaining sequence data of the sequencing template, wherein said sequence data comprises sequences of the forward strand and the reverse strand of the sequencing template; and

    determining a sequence of the template nucleic acid segment by analyzing said sequence data of the sequencing template.

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