Do I need to calibrate my instrument to detect LGC Biosearch Technologies' Pulsar® 650 dye?
The Pulsar® 650 dye is appropriate for use in the Roche LightCycler® 1.2 and 2.0 qPCR machines. The use of a two color (FAM and Pulsar 650 dye) duplex assay requires spectral calibration of the machine. Spectral calibration will decrease cross-talk between channels. This is achieved by a color compensation (CC) file loaded into your LightCycler computer. Dual-labeled BHQ® probe users have the flexibility to create this file in one of three ways:
- Download a previously-saved CC file according to your instrument model
- Generate an assay-specific CC file using the Dual-labeled BHQ probes directly or
- Generate a universal CC file using T10 calibration dyes.
Unfortunately, Molecular Beacons cannot be used directly to generate a CC file.
For instructions on how to create CC files, please download our Methods of Color Compensation on the LightCycler® Instruments_FAM/Pulsar 650 Duplexed Assays guide.
Color Compensation Files: