Detection of nucleic acid reactions on bead arrays
First Claim
1. A method of detecting a plurality of target nucleic acid sequences, comprising:
- a) providing one or more first solid supports comprising a plurality of target sequences that are immobilized to said first solid supports, wherein said first solid supports comprise a plurality of beads;
b) hybridizing a plurality of different first primers to first portions of said plurality of target sequences, wherein each of said different first primers comprises an adapter sequence exogenous to said target sequences;
c) hybridizing a plurality of different second primers to second portions of said plurality of target sequences, thereby forming a plurality of hybridization complexes;
d) extending said first or said second primers, and ligating said first and second primers together to form a plurality of different modified primers;
e) hybridizing said adapter sequences of said modified primers or their complements to an array of capture probes attached to discrete sites on a second solid support;
f) modifying said capture probes by polymerase extension, andg) detecting said modified capture probes, 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
31 Claims
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1. A method of detecting a plurality of target nucleic acid sequences, comprising:
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a) providing one or more first solid supports comprising a plurality of target sequences that are immobilized to said first solid supports, wherein said first solid supports comprise a plurality of beads; b) hybridizing a plurality of different first primers to first portions of said plurality of target sequences, wherein each of said different first primers comprises an adapter sequence exogenous to said target sequences; c) hybridizing a plurality of different second primers to second portions of said plurality of target sequences, thereby forming a plurality of hybridization complexes; d) extending said first or said second primers, and ligating said first and second primers together to form a plurality of different modified primers; e) hybridizing said adapter sequences of said modified primers or their complements to an array of capture probes attached to discrete sites on a second solid support; f) modifying said capture probes by polymerase extension, and g) detecting said modified capture probes, 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)
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Specification