Yes, Fluorescein needs to be added to the PCR or RT-PCR mix for probe-based Crystal Digital PCR™ experiments. If you wish to use EvaGreen® for amplicon detection, you need to add Alexa Fluor® 647 to the mix. For more information or queries, please contact our Support Team at firstname.lastname@example.org or email@example.com (North America) [...]
No, we do not recommend that you use ROX as a reference dye. This dye may precipitate in certain conditions in droplets, which hinders software analysis.
First, you may prepare a solution at 1mg/ml (2.6 mM) in molecular grade water, which you can then dilute to generate a 100 uM stock solution. Finally, we recommend preparing a working solution at 1 uM, which you can add directly to the PCR mix. For more information or queries, please contact our Support [...]
You need to resuspend fluorescein in molecular-grade water.
You may order fluorescein from VWR, catalog name: Fluorescein Sodium Salt, High Purity Grade, Catalog number: 0681-100 g.
Yes. You may use Alexa Fluor® 647 as a reference dye, however, you need to use another analysis configuration file, which we will provide. For more information or queries, please contact our Support Team at firstname.lastname@example.org or email@example.com (North America)
No. The Crystal Miner software needs a basal fluorescent signal to be able to count all droplets, this is why we add fluorescein to the mix.
The fluorescein is added to the PCR mix to provide a basal fluorescence signal for droplet recognition (in the blue detection channel) by the Crystal Miner software.
Fluorescein is added to the PCR/ RT-PCR mix at a final concentration of 100 nM