Compositions, Methods, and Kits for Selective Amplification of Nucleic Acids
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Abstract
The current teachings are directed to compositions, methods, and kits for selectively amplifying and for detecting target sequences. In some embodiments, a circularizable probe and/or a probe pair are disclosed for selectively amplifying target sequences. Methods for selectively amplifying target sequences are also disclosed, as are methods for detecting selectively amplified target sequences. Certain embodiments of the disclosed methods comprise a circularizable probe, a probe pair, comprising a first probe and a second probe, or both. In certain embodiments, a multiplicity of different circularizable probes, a multiplicity of different probe sets, or a multiplicity of different circularizable probes and a multiplicity of different probe sets are provided to selectively amplify or to detect a multiplicity of different target sequences, typically in a multiplex reaction. According to certain disclosed methods, surrogates of the target sequences are selectively amplified, including without limitation ligated probes, first amplification products, second amplification products, or combinations thereof. In some embodiments, selectively amplified target sequences or their surrogates are detected, directly or indirectly, indicating the presence of the corresponding target sequence. Kits to facilitate the performance of the disclosed methods are also provided.
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Citations
24 Claims
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1-9. -9. (canceled)
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10. A method for selectively amplifying a target sequence in the presence of a multiplicity of non-target sequences comprising:
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(1) hybridizing a probe with the target sequence, wherein the probe comprises three of the four nucleotide bases;
(1) A, (2) C, (3) G, and (4) at least one of T and U, but not the fourth nucleotide base; and(2) selectively amplifying the target sequence in a first reaction composition comprising; (a) an extending enzyme and (b) three of the four nucleotide bases;
(1) A, (2) C, (3) G, and (4) at least one of T and U, but not the fourth nucleotide base,to thereby generate a first amplification product, wherein each of the three nucleotide bases in the reaction composition is the complement of one of the three nucleotide bases in the probe. - View Dependent Claims (11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23)
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24-135. -135. (canceled)
Specification