Oligonucleotides containing pyrazolo 3,4-d]pyrimidines for hybridization and mismatch discrimination

(57) The oligonucleotide by which the summary singular number or two or more pudding 残 machines were replaced by ピラゾロ 3, 4*d pyrimidine shows the improved hybridization characteristic. The oligonucleotide which has the 3 and 4*ピラゾロ d pyrimidine base analog has a dissolution temperature higher than the non-replacing oligonucleotide of the same arrangement. Therefore, a higher signal is acquired in the assay containing the hybridization of an oligonucleotide probe to target polynucleotide. Furthermore, when the 3 and 4*ピラゾロ d pyrimidine content oligonucleotide is used as a hybridization probe, Inharmonious distinctiveness improves and let them be things useful as the probe and primer for a hybridization, amplification and a sequence determination procedure, and the thing for which inharmonious discernment of singleness or plurality, and a nucleotide is needed especially.