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Figure 1 | BMC Microbiology

Figure 1

From: Sensitive multiplex PCR assay to differentiate Lyme spirochetes and emerging pathogens Anaplasma phagocytophilum and Babesia microti

Figure 1

Denaturation 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.

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