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.
Claims29
A set of nucleic acid constructs, wherein the constructs of the set comprise:
- a region of interest (ROI);
- at least one unique identifier;
- and
- at least one amplification site;
- wherein the set of constructs comprises at least two constructs with the same ROI and different unique identifiers.
The set of claim 2, wherein the constructs comprise two or more amplification sites flanking the ROI and the unique identifier.
The set of claim 5, wherein the constructs comprise at least two excision sites flanking the ROI.
A method for determining the sequence of nucleic acids from a sample, comprising:
- providing nucleic acids from the sample;
- associating the individual nucleic acids with an oligonucleotide comprising a unique identifier and one or more amplification sites to create a set of tagged constructs;
- amplifying the tagged constructs so that the amplified set comprises at least two copies of each tagged construct comprising a unique identifier;
- and
- determining the sequence of at least two amplified constructs comprising the same unique identifier.
A method for identifying nucleic acids having a desired sequence, comprising:
- providing a master set of oligonucleotide constructs comprising an ROI, at least one unique identifier, and at least one amplification site;
- amplifying all or a portion of the master set;
- and
- identifying constructs having ROIs with the desired sequence by sequencing at least two constructs from the amplified set that comprise the same unique identifier;
- wherein the constructs having ROIs with the desired sequence can be identified from other constructs containing substantially the same ROI by their unique identifier.
A method for selecting nucleic acids having a desired sequence, comprising:
- providing a master set of oligonucleotide constructs wherein the constructs of the set comprise an ROI, at least one unique identifier, and at least one amplification site;
- amplifying all or a portion of the master set;
- identifying constructs having ROIs with the desired sequence by sequencing at least two constructs from the amplified set that comprise the same unique identifier;
- and
- isolating constructs comprising ROIs having the desired sequence;
- wherein the constructs having ROIs with the desired sequence can be identified from other constructs containing substantially the same ROI by their unique identifier.
Dependencies are inferred from the claim text.