Figure 1From: Sensitive multiplex PCR assay to differentiate Lyme spirochetes and emerging pathogens Anaplasma phagocytophilum and Babesia microtiDenaturation profile analysis of molecular beacon probes used in this study. Melting curves of the RecA3 molecular beacon (A) in the presence of a complementary target sequence (green line), or in the absence of any target (buffer only control, dotted line) were generated. The fluorescence intensities indicate that the molecular beacon exists either as a hybrid with its perfect complementary target sequence, exhibiting high fluorescence from 25°C to 55°C, or in its free state in the form of a stem-loop structure with fluorescence quenched in a temperature range of 25–65oC as depicted by the cartoons. At higher temperatures (more than 70oC) the molecular beacon probe denatures and exhibits high fluorescence intensities in control. Similarly, probe-target hybrid also denatures at higher temperature releasing the target and diminishing the fluorescence as the probe returns to hairpin-loop structure. A similar analysis of the BmTPK, APH1387 and ACTA1 molecular beacon probes depicted a temperature and fluorescence profile (B, C, and D), which is similar to the RecA3 molecular beacon probe.Back to article page