qPCR dye calibration standards
LGC, Biosearch Technologies offers CAL Fluor™ and Quasar™ dyes that span a wide range of emission wavelengths, for maximum flexibility in experimental design. Dye calibration standards are formulated with the fluorophore covalently attached to an oligo-dT (10-mer), to better mimic the signal from a fluorescent probe.
Calibration is required by many real-time thermal cyclers in order to generate a spectral profile for the fluorescent signal. Dye information is stored within the instrument’s software settings, and is used to control bleed-through into adjacent channels. The analysis software anticipates how much fluorescence to expect from each dye during amplification, and will subtract out signal from inappropriate filter-sets. This crosstalk is particularly consequential to a multiplexed assay, since multiple targets are amplified simultaneously but detected independently. Different fluorescent reporters are used to signal each target and so calibration with dye standards allows the instrument to resolve their overlapping spectra.
On certain instruments, all dyes that will be multiplexed together must be calibrated together, and so a FAM standard is available as well. Please inquire with firstname.lastname@example.org to discuss the custom synthesis of any dye not currently available as a calibration standard.
For multiplexed assays comprised of CAL Fluor and Quasar dyes, use dye calibration standards to fine-tune the performance of the qPCR thermal cycler.