Phosphotriester-modified oligonucleotide primers for nucleic acid amplification
First Claim
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1. A method of amplifying nucleic acids, said method comprising:
- amplifying nucleic acid using a modified oligonucleotide primer,wherein said modified oligonucleotide primer comprises a modification group at one or two internucleotide linkages;
wherein said modification groups are at the n, n-1, n-2, or n-3 positions, wherein n is the 3′
terminal internucleotide linkage;
wherein the presence of said modification group impairs nucleic acid polymerase extension of said modified oligonucleotide primer;
wherein said modification group can be non-reversibly thermally dissociated; and
wherein said modified oligonucleotide primer has structure I as follows
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Abstract
The present invention provides methods and compositions for nucleic acid amplification. These methods involve the use of oligonucleotide primers in temperature dependent nucleic acid amplification reactions. In certain aspects, the methods are accomplished by use of certain modified oligonucleotide primers which provide utility in nucleic acid amplification. In preferred embodiments, the oligonucleotide primers are modified with particular chemical groups such as esters.
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34 Claims
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1. A method of amplifying nucleic acids, said method comprising:
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amplifying nucleic acid using a modified oligonucleotide primer, wherein said modified oligonucleotide primer comprises a modification group at one or two internucleotide linkages; wherein said modification groups are at the n, n-1, n-2, or n-3 positions, wherein n is the 3′
terminal internucleotide linkage;wherein the presence of said modification group impairs nucleic acid polymerase extension of said modified oligonucleotide primer; wherein said modification group can be non-reversibly thermally dissociated; and wherein said modified oligonucleotide primer has structure I as follows - 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, 31, 32, 33, 34)
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Specification