PRIMER FOR NUCLEIC ACID DETECTION
First Claim
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1. A method of making a labeled sequence-specific primer, comprising:
- adding a universal nucleic acid sequence to a sequence-specific primer, thereby generating a labeled sequence-specific primer,wherein the universal nucleic acid sequence comprisesa 5′
end,a 3′
end; and
a labeled nucleotide flanked on each side by at least one nucleotide, wherein nucleotides complementary to the nucleotides flanking the labeled nucleotide quench a detectable signal from the label when the labeled sequence-specific primer is hybridized with its complementary nucleic acid molecule and the complementary nucleic acid is synthesized,wherein the sequence-specific primer can hybridize to a target nucleic acid sequence, andwherein the universal nucleic acid sequence does not substantially hybridize to the target nucleic acid sequence recognized by the sequence-specific primer.
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Abstract
This application provides universal labeled primers for detection and amplification of nucleic acid molecules. These universal primers can be attached to the 5′-end of a target sequence-specific primer. In particular examples, the universal primer includes a labeled nucleotide flanked on both sides a nucleotide whose complement nucleotides changes a detectable signal from the label when the universal primer hybridizes with its complementary nucleic acid molecule. Also disclosed are methods of using the universal primer in nucleic acid amplification, such as real-time PCR.
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Citations
30 Claims
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1. A method of making a labeled sequence-specific primer, comprising:
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adding a universal nucleic acid sequence to a sequence-specific primer, thereby generating a labeled sequence-specific primer, wherein the universal nucleic acid sequence comprises a 5′
end,a 3′
end; anda labeled nucleotide flanked on each side by at least one nucleotide, wherein nucleotides complementary to the nucleotides flanking the labeled nucleotide quench a detectable signal from the label when the labeled sequence-specific primer is hybridized with its complementary nucleic acid molecule and the complementary nucleic acid is synthesized, wherein the sequence-specific primer can hybridize to a target nucleic acid sequence, and wherein the universal nucleic acid sequence does not substantially hybridize to the target nucleic acid sequence recognized by the sequence-specific primer. - 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)
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Specification