Multiplexed analytical platform
The invention provides a system and reagents for carrying out multiplexed analytical measurements using the hybridization of oligonucleotide tags to addressable solid phase supports to obtain simultaneous readouts from hundreds to tens of thousands of reactions. In one aspect of the invention, analyte interaction moieties, such as oligonucleotide probes, are produced that are each labeled with a unique synthesis tag constructed from a set of two-nucleotide “words.” During or after an assay, the synthesis tags are converted to hybridization tags that are used in the readout step. Hybridization tags are constructed to maximize discrimination in the readout. Hybridization tags of the invention are preferably constructed from oligonucleotide “words” selected from a minimally cross-hybridizing set using combinatorial synthesis techniques. In a further aspect, discrimination in the readout is enhanced by employing hybridization tags that are constructed with words having the “comma-less” property and by minimizing or eliminating extraneous nucleic acids associated with the hybridization tag during a readout step.
1. A system for detecting one or more properties of each of a plurality of analytes, the system comprising:
a first set of analyte-interaction nucleic acid moieties capable of reacting with the plurality of analytes under assay reaction conditions, each analyte-interaction nucleic acid moiety having a synthesis tag comprising a concatenation of words;
conversion reagents for converting each synthesis tag into a hybridization tag and for releasing the hybridization tag from the analyte-interaction nucleic acid moieties, wherein the hybridization tag comprises one or more words determined by a corresponding synthesis tag, each word of the hybridization tag being longer than each word of its corresponding synthesis tag; and
a solid phase detection array for detecting the released hybridization tags, thereby detecting the one or more properties of each of the plurality of analytes.
2. The system of claim 1 wherein each of said words of said hybridization tag is selected from a minimally cross-hybridizing set, each word of the minimally cross-hybridizing set being an oligonucleotide having a length in the range of from three to nine nucleotides and differing in nucleotide sequence from every other word of such set by at least two nucleotides.
3. The system of claim 2 wherein said hybridization tag is comma-less.
4. The system of claim 3 wherein said analyte-interaction moiety is an oligonucleotide.
5. The system of claim 4 wherein said words of said hybridization tag each have a length in the range of from 4 to 5 nucleotides, wherein said words of said synthesis tag have a length of two nucleotides, and wherein said solid phase detection array is a microarray having from 100 to 100,000 hybridization sites.
6. A solid phase support having attached a plurality of tag complements, the solid phase support comprising:
a plurality of addressable sites, each addressable site having copies of only one tag complement, the tag complement having the structure:
W 1 W 2 . . . W n
wherein each W i for i=1, 2, . . . n is selected from a group of oligonucleotides having nucleotide sequences:
AICAT
GTCTA
GAACT
TGTCA
CTTGT
ACAGA
TCITT
CAGAA
or
CTGTA
CAAGT
TCTGA
ACIAT
AGACA
TICTT
GACAA
GTTCT.