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US 10,501,793 B2Granted
Nucleic acid constructs and methods of use
AssigneePrognosys Biosciences, Inc.
InventorMark S. Chee
Life of this patent
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Abstract
The present invention provides oligonucleotide constructs, sets of such oligonucleotide constructs, and methods of using such oligonucleotide constructs to provide validated sequences or sets of validated sequences corresponding to desired ROIs. Such validated ROIs and constructs containing these have a wide variety of uses, including in synthetic biology, quantitative nucleic acid analysis, polymorphism and/or mutation screening, and the like.
Claims21
A method to analyze nucleic acid sequences, comprising:
- attaching a unique identifier nucleic acid sequence (UID) from a pool of UIDs to a first end of each strand of a plurality of analyte nucleic acid fragments to form a plurality of uniquely identified analyte nucleic acid fragments wherein the pool of UIDs is in excess of the plurality of analyte nucleic acid fragments;
- redundantly determining nucleotide sequence of a uniquely identified analyte nucleic acid fragment, wherein determined nucleotide sequences which share a UID form a family of members;
- comparing determined nucleotide sequences between members of a family;
- and
- identifying a nucleotide sequence as accurately representing an analyte nucleic acid fragment when the sequence is a consensus sequence of the determined nucleotide sequences of the members of the family and wherein the family contains at least 2 members.
2 more elements
A method to analyze nucleic acid sequences, comprising:
- attaching a unique identifier nucleic acid sequence (UID) of about 10 to about 30 bases from a pool of UIDs to a first end of each strand of a plurality of analyte nucleic acid fragments using at least two cycles of polymerase chain reaction to form uniquely identified analyte nucleic acid fragments, wherein the pool of UIDs is in excess of the plurality of analyte nucleic acid fragments;
- redundantly determining nucleotide sequence of a uniquely identified analyte nucleic acid fragment, wherein determined nucleotide sequences which share a UID form a family of members;
- comparing determined nucleotide sequences between members of a family;
- identifying a nucleotide sequence as accurately representing an analyte nucleic acid fragment when the sequence is a consensus sequence of the determined nucleotide sequences of the members of the family and wherein the family contains at least 2 members;
- and
- comparing the nucleotide sequence identified as accurately representing an analyte nucleic acid fragment to a reference sequence and identifying a single base substitution, insertion, or deletion in the nucleotide sequence relative to the reference sequence.
3 more elements
Dependencies are inferred from the claim text.