New cyanine dyes as base surrogates in PNA: forced intercalation probes (FIT-probes) for homogeneous SNP detection.

摘要: Abstract Forced intercalation probes (FIT-probes) are nucleic acid probes, in which an intercalator cyanine dye such as thiazole orange (TO) serves a replacement of canonical nucleobase. These signal hybridization by showing strong increases fluorescence. TO FIT-probes responds to adjacent base mismatches attenuation fluorescence intensities at conditions where both matched and mismatched target DNA bound. The interesting features labeled posed the question whether forced concept can be extended other dyes family. Herein, we present synthesis three asymmetrical their incorporation into PNA-conjugates means divergent linear solid-phase synthesis. Melting analysis revealed that affinity PNA remained high irrespective nucleobase cyanines YO (oxazole yellow), MO or JO. Of new tested dye–PNA-conjugates, YO-containing has properties useful for homogeneous SNP detection. YO–PNA is demonstrated presence fully complementary up 20-fold enhancement In addition, emission discriminates against single Oxazole yellow (YO) surrogate may prove multiplex detection mutations non-stringent conditions.