Control for Primer Dimers in Probe based qPCR assays
- Are you using probe based qPCR assays and would need a dissociation curve?
- Are you performing gel electrophoresis after your qPCR to control your probe based qPCR run, risking contamination?
- Do you need a non-specific reporter to detect any amplified dsDNA?
In quantitative real-time (qPCR), the detection with unspecific dyes includes artefacts such as primer dimers, making it beneficial in some cases to use sequence specific probes. But when using probes, dissociation curves cannot be performed to study what has been amplified in each reaction. The option is to use gel electrophoresis for troubleshooting, leading to extra workload and extremely high risks of contamination.
Best of both worlds
BOXTO shows a strong fluorescence increase when bound to dsDNA and can be detected on the JOE channel. Use as an unspecific dye for qPCR and in combination with any FAM-labeled probe.
TATAA Biocenter has developed BOXTO, an unsymmetric cyanine dye for use in qPCR applications. BOXTO can be used as an unspecific dye for qPCR applications or other applications where staining of dsDNA is wanted. The dye has absorbance and emission wavelengths that can be detected on the JOE channel on most common real-time PCR instrumentation and shows a strong fluorescence increase when bound to dsDNA. The dye can be used in combination with any FAM-labeled probe and a dissociation curve can be performed after amplification. The quantification is performed on the FAM-channel and the dissociation curve on the JOE-channel. The dye is an excellent control for primer dimers in probe based qPCR assays.