Method for analyzing a nucleic acid sequence
AssigneeSysmex Corporation
InventorFrank Holtrup
Life of this patent
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Abstract
Disclosed is a method for sequencing a nucleotide sequence of a target nucleic acid. The method comprises: providing a pool of amplicons, wherein the pool of amplicons is prepared by attaching a unique identifier to the target nucleic acid, and amplifying by PCR the target nucleic acid to which the unique identifier is attached; and sequencing the amplicons comprising the unique identifier and the target nucleic acid. In the method, a nucleotide sequence of the unique identifier comprises both a random nucleotide (N) and a predetermined nucleotide.
Claims12
A method for sequencing a nucleotide sequence of a target nucleic acid, comprising:
- providing a pool of amplicons, wherein the pool of amplicons is prepared by a process comprising PCR amplification of a target nucleic acid using primers each containing a unique identifier;
- and
- sequencing the amplicons comprising the unique identifier and the target nucleic acid,
- wherein the nucleotide sequence of the unique identifier contains a plurality of random nucleotides (N) and a plurality of predetermined nucleotides,
- wherein a nucleotide at the 5′ end of the unique identifier is a random nucleotide (N), and a nucleotide at the 3′ end of the unique identifier is a random nucleotide (N), and
- wherein all of said predetermined nucleotides are each independently selected from the group consisting of nucleotides comprising adenine as a base, and nucleotides comprising thymine as a base.
A method for sequencing a nucleotide sequence of a target nucleic acid, comprising:
- providing a pool of amplicons, wherein the pool of amplicons is prepared by amplifying the target nucleic acid using tagged primers each containing:
- (i) a universal primer section;
- and (ii) a tag section containing a unique identifier to produce tagged target nucleic acids, and amplifying the tagged target nucleic acids using universal primers to produce the pool of amplicons;
- and
- sequencing the amplicons comprising the unique identifier and the target nucleic acid,
- wherein the nucleotide sequence of the unique identifier contains a plurality of random nucleotides (N) and a plurality of predetermined nucleotides,
- wherein a nucleotide at the 5′ end of the unique identifier is a random nucleotide (N), and a nucleotide at the 3′ end of the unique identifier is a random nucleotide (N), and
- wherein all of said predetermined nucleotides are each independently selected from the group consisting of nucleotides comprising adenine as a base, and nucleotides comprising thymine as a base.
Dependencies are inferred from the claim text.