Compositions and methods for nucleic acid sequencing

CAFC
  • US 9,404,146 B2
  • Filed: 04/19/2013
  • Issued: 08/02/2016
  • Est. Priority Date: 03/28/2008
  • Status: Active Grant
First Claim
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1. A method of sequencing nucleic acid molecules with improved accuracy, comprising:

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

    linking a 3′

    end of the forward strand to a 5′

    end of the reverse strand of each of said nucleic acid molecules, thereby forming a plurality of template molecules, wherein said 3′

    end of the forward strand and said 5′

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

    performing at least one single-molecule sequencing process on each of said plurality of template molecules, wherein the forward strand and the reverse strand in each of said plurality of template molecules are fully complementary to each other, each of said at least one single-molecule sequencing process consisting of sequencing only one single template molecule of said plurality of template molecules, thereby generating a sequence read from each of said plurality of template molecules, said sequence read comprising sequences of both the forward strand and the reverse strand of each of said plurality of template molecules; and

    determining the sequence of each of said plurality of template molecules by analyzing said sequence read from each of said plurality of template molecules, including comparing the sequences of the forward strand and the reverse strand from each of said plurality of template molecules, thereby improving sequencing accuracy of said nucleic acid molecules.

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