CHEMICALLY 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 more internucleotide linkages;
wherein said modification group can be non-reversibly thermally dissociated; and
wherein said modified oligonucleotide primer has structure I as follows wherein;
Nuc is a nucleoside within the primer sequence;
U and Z are independently O, S, Se, NR9, or CR9R10;
R9 and R10 are each independently hydrogen or optionally substituted straight or branched hydrocarbyl having from 1-20 carbon atoms, wherein each may independently include at least one substituent selected from halo, oxo, hydroxyl, alkoxy, aryloxy, amino, amido or a detectable label;
Y is O, S or Se;
W is O, S, S(O), S(O)2, Se, C(O), C(S), C(O)NH, NH or NR9; and
Q is a modification group comprising one or more thermally cleavable groups.
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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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Citations
39 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 more internucleotide linkages;
wherein said modification group can be non-reversibly thermally dissociated; and
wherein said modified oligonucleotide primer has structure I as follows wherein;
Nuc is a nucleoside within the primer sequence;
U and Z are independently O, S, Se, NR9, or CR9R10;
R9 and R10 are each independently hydrogen or optionally substituted straight or branched hydrocarbyl having from 1-20 carbon atoms, wherein each may independently include at least one substituent selected from halo, oxo, hydroxyl, alkoxy, aryloxy, amino, amido or a detectable label;
Y is O, S or Se;
W is O, S, S(O), S(O)2, Se, C(O), C(S), C(O)NH, NH or NR9; and
Q is a modification group comprising one or more thermally cleavable groups. - 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, 27, 28, 29, 30, 31, 32, 34, 35, 36, 37, 38, 39)
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24. An oligonucleotide primer for nucleic acid amplification comprising:
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a nucleic acid sequence;
wherein said nucleic acid sequence has one or more modification groups; and
wherein said modification group comprises one or more thermally cleavable groups. - View Dependent Claims (25)
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26. A kit for nucleic acid amplification comprising one or more modified oligonucleotides wherein said modified oligonucleotides comprises one or more thermally cleavable groups.
Specification