Nucleic acid amplification using a reversibly modified oligonucleotide
First Claim
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1. A method for regenerating a 3′
- hydroxyl group, the method comprising;
using a mixture comprising at least one reversibly modified oligonucleotide having a non-hydroxyl group 3′
end which can be converted into a 3′
hydroxyl group, wherein the oligonucleotide has a carboxylic acid ester group at its 3′
end;
exposing the mixture to a first chemical and a first range of temperature, wherein the first chemical and the first range of temperature regenerate the 3′
hydroxyl group; and
regenerating the 3′
hydroxyl group of the at least one reversibly modified oligonucleotide having a non-hydroxyl 3′
end.
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Abstract
The present invention provides a method for amplification of a target nucleic acid sequence or signal, wherein an amplification reaction mixture is used which contains at least one reversibly modified oligonucleotide having a non-hydroxyl group 3′ end which can be converted into a hydroxyl 3′ end upon exposure to a chemical and/or irradiation and/or a range of temperature. The present invention also provides a reversibly modified oligonucleotide as described above, and a nucleic acid amplification reaction mixture and kit comprising such an oligonucleotide.
42 Citations
25 Claims
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1. A method for regenerating a 3′
- hydroxyl group, the method comprising;
using a mixture comprising at least one reversibly modified oligonucleotide having a non-hydroxyl group 3′
end which can be converted into a 3′
hydroxyl group, wherein the oligonucleotide has a carboxylic acid ester group at its 3′
end;exposing the mixture to a first chemical and a first range of temperature, wherein the first chemical and the first range of temperature regenerate the 3′
hydroxyl group; andregenerating the 3′
hydroxyl group of the at least one reversibly modified oligonucleotide having a non-hydroxyl 3′
end. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 18)
- hydroxyl group, the method comprising;
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9. A method for regenerating a 3′
- hydroxyl group, the method comprising;
using a mixture comprising at least one reversibly modified oligonucleotide having a non-hydroxyl group 3′
end which can be converted into a 3′
hydroxyl group, wherein the at least one reversibly modified oligonucleotide has a silyl ether group at its 3′
end;exposing the mixture to a first chemical and a first range of temperature, wherein the first chemical and the first range of temperature regenerate the 3′
hydroxyl group; andregenerating the 3′
hydroxyl group of the at least one reversibly modified oligonucleotide having a non-hydroxyl 3′
end. - View Dependent Claims (10, 19, 21)
- hydroxyl group, the method comprising;
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11. A method for regenerating a 3′
- hydroxyl group, the method comprising;
using a mixture comprising at least one reversibly modified oligonucleotide having a non-hydroxyl group 3′
end which can be converted into a 3′
hydroxyl group, wherein the at least one reversibly modified oligonucleotide has an ether group at its 3′
end;exposing the mixture to a first chemical and a first range of temperature, wherein the first chemical and the first range of temperature regenerate the 3′
hydroxyl group; andregenerating the 3′
hydroxyl group of the at least one reversibly modified oligonucleotide having a non-hydroxyl 3′
end. - View Dependent Claims (12, 13, 14, 15, 16, 17, 20, 22)
- hydroxyl group, the method comprising;
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23. A method for regenerating 3′
- hydroxyl groups, the method comprising;
using a mixture comprising at least two reversibly modified oligonucleotides, wherein each oligonucleotide has a different non-hydroxyl group 3′
end which can be converted into 3′
hydroxyl group, wherein the non-hydroxyl group is a carboxylic acid ester, a silyl ether, or an ether, andexposing the mixture to two different chemicals and at least one range of temperature, wherein the chemicals and the range of temperature regenerate the 3′
hydroxyl groups. - View Dependent Claims (24, 25)
- hydroxyl groups, the method comprising;
Specification