Colour compensation in LightCycler 480

The LightCycler 480 instrument requires color compensation for a 4-plex assay, as outlined in its manual. Color compensation can be applied using a file from a different assay that is using FAM, HEX, Texas Red (ROX), and Cy5.

In case no color compensation file is available, RTPCR010 can be assayed with some specifics. The instrument can correctly read FAM, HEX, and ROX, however, Cy5 would be positive for any sample with N gene amplification due to cross-talk between ROX and Cy5. Since Cy5 is used as an internal control in RTPCR010 and should be positive in all samples, RTPCR010 could be used, but ideally color compensation would resolve this.

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  In attached files.