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Labelling Overview - Dual Labelled Probes
| Fluorescence | Fluorescein (FAM) quenched by BHQ1 quencher | | Application | Set the cycler to FAM channel and/or set excitation wavelength to 494 nm and emission wavelength to 525 nm |
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| Fluorescence | Hexachlorofluorescein (HEX) quenched by BHQ1 quencher | | Application | Set the cycler to JOE channel and/or set excitation wavelength to 530 nm and emission wavelength to 556 nm |
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| Fluorescence | Cyanide dye Cy3 (ex 550 nm/em 570 nm) quenched by BHQ2 quencher | | Application | Set the cycler to Cy3 channel and/or set excitation wavelength to 470 nm (FAM) or better to 530 nm and emission wavelength to filter bandpass 570 nm or highpass 585 nm (585hp for the cycler RotorGene, Corbett Research) |
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| Fluorescence | Cyanide dye Cy5 (ex 649 nm/em 670 nm) quenched by BHQ2 quencher | | Application | Set the cycler to Cy5 channel and/or LC-Red640 channel or set excitation wavelength to 640 nm (625 nm for the cycler RotorGene, Corbett Research) and emission wavelength to filter bandpass 660 nm |
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| Fluorescence | Ruthenium dye PULSAR (ex 470 nm/em 650 nm) quenched by BHQ2 quencher | | Application | PULSAR is designed especially for multiplex experiments in LightCycler 1.2. As for multiplex analyses in LightCycler, F1 channel should be set for FAM probe and F3 channel for BHQ2-PULSAR probe. In other cyclers (for example, Corbett Research - RotorGene), a new channel must be form with excitation at 470 nm and emission at 650 nm, i.e. 610 nm high pass or 660 nm high pass (610hp or 660hp). Do not use 610bp! |
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