Supplements

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Thermal Cycler Color Compensation Kit

Applications

LightCycler^®480 I or II

Contents

The Calibration dyes contain labeled oligonucleotides for the determination of crosstalk corresponding to each individual dye resp. channel being used in the multiplex assay, and a blank reaction buffer without dye for the determination of background fluorescence of the buffer.

Performing a Color Compensation Experiment

Although the filter sets in Multiplex PCR cyclers are optimized for the commonly used fluorescence dyes used in PCR experiments, some crosstalk between the channels might be seen. Performing a Color Compensation Experiment will correct the crosstalk to allow proper interpretation of a multiplex PCR run.

The Color Compensation reagents are designed for the generation of color compensation objects on the LightCycler^®480 I or II system to allow interpretation of multiplex experiments. Subsequently, the generated file may be used for analyzing runs with our multiplex PCR kit Onar^®MRSA Detection Kit or other kits using the same type of dyes.

As real-time cyclers may vary in their fluorescence detection sensitivity, color compensation experiments must be performed on each specific multiplex-PCR instrument.

In the case of overcompensation or remaining crosstalk it is recommended to adjust the Calibration dye concentration used for performing the color compensation experiment to result in an equal level of (absolute) fluorescence signal.