DNA amplification technology
First Claim
Patent Images
1. A nucleic acid complex, comprising a double-stranded target nucleic acid and a first oligonucleotide primer and a second oligonucleotide primer hybridized thereto,wherein the first oligonucleotide primer comprises a 3′
- end having a sequence substantially complementary to a binding site on the first strand of the target nucleic acid and a 5′
end substantially complementary to a binding site on the second strand of the target nucleic acid opposite to the binding she of the 3′
end of the first oligonucleotide primer;
wherein the second oligonucleotide primer comprises a 3′
end having a sequence substantially complementary to a binding site on the second strand of the target nucleic acid and a 5′
end substantially complementary to a binding site on the first strand of the target nucleic acid opposite to the binding site of the 3′
end of the second oligonucleotide primer;
wherein on the first strand, the binding site of the 3′
end of the first oligonucleotide primer is 3′
to the binding site of the 5′
end of the second oligonucleotide primer; and
wherein on the second strand, the binding site of the 3′
end of the second oligonucleotide primer is 3′
to the binding site of the 5′
end of the first oligonucleotide primer.
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Abstract
Methods and reagents suitable for conducing polymerase chain reaction are described. In particular, a nucleic acid amplification design strategy and thermal cycling profile to enable efficient amplification of multiple nucleic acid targets along with improved sensitivity is disclosed. The present disclosure also describes methods and devices for increasing the melting temperature (Tm) of a primer.
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Citations
24 Claims
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1. A nucleic acid complex, comprising a double-stranded target nucleic acid and a first oligonucleotide primer and a second oligonucleotide primer hybridized thereto,
wherein the first oligonucleotide primer comprises a 3′ - end having a sequence substantially complementary to a binding site on the first strand of the target nucleic acid and a 5′
end substantially complementary to a binding site on the second strand of the target nucleic acid opposite to the binding she of the 3′
end of the first oligonucleotide primer;wherein the second oligonucleotide primer comprises a 3′
end having a sequence substantially complementary to a binding site on the second strand of the target nucleic acid and a 5′
end substantially complementary to a binding site on the first strand of the target nucleic acid opposite to the binding site of the 3′
end of the second oligonucleotide primer;wherein on the first strand, the binding site of the 3′
end of the first oligonucleotide primer is 3′
to the binding site of the 5′
end of the second oligonucleotide primer; andwherein on the second strand, the binding site of the 3′
end of the second oligonucleotide primer is 3′
to the binding site of the 5′
end of the first oligonucleotide 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)
- end having a sequence substantially complementary to a binding site on the first strand of the target nucleic acid and a 5′
Specification