BOXTO

A new dye for qPCR

BOXTO

BOXTO is a derivative of BEBO, an unsymmetric cyanine dye developed by TATAA Biocenter for use in qPCR applications. The dye has absorbance and emission wavelengths that can be detected on the JOE channel on most common real-time PCR instruments, and shows a very strong fluorescence increase when bound to dsDNA. BOXTO can be used as an unspecific dye for real-time PCR applications or other applications where staining of dsDNA is wanted. The BEBO-family of dyes is patent pending.

Best of both worlds?

Since unspecific dyes also detect artefacts such as primerdimers, in some cases using sequence specific probes, may be beneficial. On the other hand, when using probes dissociation curves cannot be performed to study what has been amplified in each reaction, and in some cases gel electrophoresis has to be performed to troubleshoot leading to extremely high risk of contamination. BOXTO can be used in combination with any FAM-labelled probe and a dissociation curve can be performed after amplification. Quantification is then performed on the FAM-channel and dissociation curve on the JOE-channel.

Figure: Using BOXTO in combination with a molecular beacon FAM-labelled probe. Detecting on the FAM-channel does not detect the NTC containing only primerdimers (red curves). These are detected in the JOE-channel with BOXTO (inset) in the same reaction tube. Dissociation curve analysis can be performed after amplifi cation using BOXTO on the JOE-channel. Here 0,7µM of BOXTO was used in combination with 0,4µM of the molecular beacon probe.

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