Modified oligonucleotides for mismatch discrimination
First Claim
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1. A method for primer extension, the method comprising incubating a polynucleotide containing a target sequence with one or more oligonucleotide primers complementary to the target sequence, in the presence of a polymerizing enzyme and nucleotide substrates under conditions favorable for polymerization;
- wherein at least one of the oligonucleotide primers contains a modified base selected from the group consisting of an unsubstituted pyrazolo[3,4-d]pyrimidine, a 3-substituted pyrazolo[3,4-d]pyrimidine and a 5-substituted pyrimidine base, in place of a purine or pyrimidine base.
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Abstract
Modified oligonucleotides are provided containing bases selected from unsubstituted and 3-substituted pyrazolo[3,4-d]pyrimidines and 5-substituted pyrimidines, and optionally have attached minor groove binders and reporter groups.
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Citations
9 Claims
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1. A method for primer extension, the method comprising incubating a polynucleotide containing a target sequence with one or more oligonucleotide primers complementary to the target sequence, in the presence of a polymerizing enzyme and nucleotide substrates under conditions favorable for polymerization;
- wherein at least one of the oligonucleotide primers contains a modified base selected from the group consisting of an unsubstituted pyrazolo[3,4-d]pyrimidine, a 3-substituted pyrazolo[3,4-d]pyrimidine and a 5-substituted pyrimidine base, in place of a purine or pyrimidine base.
- View Dependent Claims (2, 3, 4, 5, 6)
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7. A method for determining the nucleotide sequence of a polynucleotide, the method comprising:
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(a) incubating the polynucleotide with a modified oligonucleotide array under hybridization conditions; and
(b) determining to which of the modified oligonucleotides in the array the polynucleotide hybridizes;
wherein a plurality of the modified oligonucleotides comprise at least one 3-substituted pyrazolo[3,4]pyrimidine in place of a purine base. - View Dependent Claims (8, 9)
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Specification