Hi,
We have Lightcycler 2.0. for RRT-PCR in our laboratory. Based on our two previous experiments; LightCycler 2.0. cannot detect MGB probe ( were labelled with FAM and VIC). What will happen If labelling the same probe sequences with FAM and TAMRA ? Does it work again? Or any TM changes ?
Many thanks for your answers in advance
Have you verified that the probe is functional on another platform? a FAM probe is a FAM probe and if properly designed should work fine on a LightCycler.
I would certainly check your design and verify that your assay is functioning properly ( by gel electrophoresis), and that your target sequence is correct (MGB are sensitive to mismatches).
P.S. "MGB" is not a probe type, it is an oligo modification that enhances hybridization - I assume you mean a TaqMan Hydrolysis probe.
MGB is not a probe, so it is not emitted fluorescence. Select "NONE" option in the equipment . The MGB increases the annealing temperature (TM) probe. So, if you use the TAMRA, your TM will change.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Molecular Biological Techniques