Detection of nucleic acid reactions on bead arrays
First Claim
1. A method of detecting a plurality of target deoxyribonucleic acids, comprising:
- a) providing a plurality of target deoxyribonucleic acids;
b) hybridizing a plurality of first primers to first portions of each of said plurality of target deoxyribonucleic acids, wherein each of said first primers comprises an adapter sequence exogenous to sequences of said target deoxyribonucleic acids, wherein said plurality of first primers are different from each other;
c) hybridizing a plurality of second primers to second portions of each of said plurality of target deoxyribonucleic acids, wherein said plurality of second primers are different from each other;
d) extending said first or said second primers, and then ligating said first and second primers together to form a plurality of modified primers comprising said adapter sequences;
e) hybridizing said adapter sequences of said modified primers to an array of capture probes attached to a solid support, thereby forming hybridized capture probes;
f) modifying said hybridized capture probes by polymerase extension to form modified capture probes, andg) detecting said modified capture probes, thereby detecting said plurality of target deoxyribonucleic acids,wherein unextended first or second primers are removed from said modified primers prior to (e).
2 Assignments
0 Petitions
Accused Products
Abstract
The present invention is directed to methods and compositions for the use of micro sphere 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.
-
Citations
26 Claims
-
1. A method of detecting a plurality of target deoxyribonucleic acids, comprising:
-
a) providing a plurality of target deoxyribonucleic acids; b) hybridizing a plurality of first primers to first portions of each of said plurality of target deoxyribonucleic acids, wherein each of said first primers comprises an adapter sequence exogenous to sequences of said target deoxyribonucleic acids, wherein said plurality of first primers are different from each other; c) hybridizing a plurality of second primers to second portions of each of said plurality of target deoxyribonucleic acids, wherein said plurality of second primers are different from each other; d) extending said first or said second primers, and then ligating said first and second primers together to form a plurality of modified primers comprising said adapter sequences; e) hybridizing said adapter sequences of said modified primers to an array of capture probes attached to a solid support, thereby forming hybridized capture probes; f) modifying said hybridized capture probes by polymerase extension to form modified capture probes, and g) detecting said modified capture probes, thereby detecting said plurality of target deoxyribonucleic acids, wherein unextended first or second primers are removed from said modified primers prior to (e). - 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)
-
Specification