OLIGONUCLEOTIDES CONTAINING PYRAZOLO[3,4-D] PYRIMIDINES FOR HYBRIDIZATION AND MISMATCH DISCRIMINATION
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Abstract
Oligonucleotides in which one or more purine residues are substituted by pyrazolo[3,4-d]pyrimidines exhibit improved hybridization properties. Oligonucleotides containing pyrazolo[3,4-d]pyrimidine base analogues have higher melting temperatures than unsubstituted oligonucleotides of identical sequence. Thus, in assays involving hybridization of an oligonucleotide probe to a target polynucleotide sequence, higher signals are obtained. In addition, mismatch discrimination is enhanced when pyrazolo[3,4-d]pyrimidine-containing oligonucleotides are used as hybridization probes, making them useful as probes and primers for hybridization, amplification and sequencing procedures, particularly those in which single- or multiple-nucleotide mismatch discrimination is required.
22 Citations
62 Claims
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1-47. -47. (canceled)
- 48. An oligonucleotide probe capable of hybridizing with a target sequence comprising a modified target binding sequence wherein one or more purine residues of said target binding sequence are substituted by a pyrazolo[3,4-d]pyrimidine residue, and further comprising an attached minor groove binder, said modified target binding sequence corresponding to and capable of binding with a target sequence containing no pyrazolo[3,4-d]pyrimidine residues.
Specification