DETECTION OF NUCLEIC ACID REACTIONS ON BEAD ARRAYS
First Claim
1. A method of detecting a plurality of target nucleic acid sequences, comprising:
- a) hybridizing a plurality of different first primers to first portions of a plurality of target sequences that are immobilized on one or more solid phase surfaces, wherein each of said different first primers comprises a different adapter sequence exogenous to said target sequences;
b) hybridizing a plurality of different second primers to second portions of said plurality of target sequences, thereby forming a plurality of hybridization complexesc) extending said first or said second primers, and ligating said first and second primers together to form a plurality of different modified primers;
d) hybridizing said adapter sequences of said modified primers or their complements to an array of capture probes attached to a second solid support; and
e) detecting the presence of said modified primers, thereby detecting said plurality of target nucleic acid sequences.
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Abstract
The present invention is directed to methods and compositions for the use of microsphere arrays to detect and quantify a number of nucleic acid reactions. The invention finds use in genotyping, i.e. the determination of the sequence of nucleic acids, particularly alterations such as nucleotide substitutions (mismatches) and single nucleotide polymorphisms (SNPs). Similarly, the invention finds use in the detection and quantification of a nucleic acid target using a variety of amplification techniques, including both signal amplification and target amplification. The methods and compositions of the invention can be used in nucleic acid sequencing reactions as well. All applications can include the use of adapter sequences to allow for universal arrays.
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Citations
34 Claims
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1. A method of detecting a plurality of target nucleic acid sequences, comprising:
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a) hybridizing a plurality of different first primers to first portions of a plurality of target sequences that are immobilized on one or more solid phase surfaces, wherein each of said different first primers comprises a different adapter sequence exogenous to said target sequences; b) hybridizing a plurality of different second primers to second portions of said plurality of target sequences, thereby forming a plurality of hybridization complexes c) extending said first or said second primers, and ligating said first and second primers together to form a plurality of different modified primers; d) hybridizing said adapter sequences of said modified primers or their complements to an array of capture probes attached to a second solid support; and e) detecting the presence of said modified primers, thereby detecting said plurality of target nucleic acid sequences. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34)
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Specification