Methods and compositions for tagging and identifying polynucleotides
First Claim
1. A composition of nucleic acid molecules made by the following method:
- ligating a discriminatory oligonucleotide tag to fragmented genomic DNA so that substantially every target polynucleotide in said fragmented genomic DNA receives a tag that comprises a unique nucleotide sequence, to produce a tagged sample; and
amplifying the target polynucleotides in said tagged sample by polymerase chain reaction to produce a plurality of amplicons that each comprise a unique tag.
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Abstract
The invention provides methods and compositions for attaching oligonucleotide tags to polynucleotides for the purpose of carrying out analytical assays in parallel and for decoding the oligonucleotide tags of polynucleotides selected in such assays. Words, or subunits, of oligonucleotide tags index submixtures in successively more complex sets of submixtures (referred to herein as “tiers” of submixtures) that a polynucleotide goes through while successive words are added to a growing tag. By identifying each word of an oligonucleotide tag, a series of submixtures is identified including the first submixture that contains only a single polynucleotide, thereby providing the identity of the selected polynucleotide. The analysis of the words of an oligonucleotide tag can be carried out in parallel, e.g. by specific hybridization of the oligonucleotide tag to its tag complement on an addressable array; or such analysis can be carried out serially by successive specific hybridizations of labeled word complements, or the like.
140 Citations
3 Claims
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1. A composition of nucleic acid molecules made by the following method:
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ligating a discriminatory oligonucleotide tag to fragmented genomic DNA so that substantially every target polynucleotide in said fragmented genomic DNA receives a tag that comprises a unique nucleotide sequence, to produce a tagged sample; and amplifying the target polynucleotides in said tagged sample by polymerase chain reaction to produce a plurality of amplicons that each comprise a unique tag. - View Dependent Claims (2, 3)
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Specification