EvaGreen® dye is spectrally similar to FAM or SYBR® Green I, which means no change in optical settings is required for using an EvaGreen®-based master mix. However, there are several differences between the dyes.
Lower background fluorescence: EvaGreen® dye has less background than SYBR® Green I due to its novel “release-on-demand” DNA-binding mechanism.
Brighter and non-inhibitory: EvaGreen® is less inhibitory in the PCR reaction than SYBR® Green, permitting the use of saturating dye concentration for maximal signal and better high-resolution DNA melt analysis.
Safer and more environmentally friendly: EvaGreen® dye is the first and only PCR dye to date designed to be environmentally safe.
Dye stability: EvaGreen® dye is very stable both during storage and under PCR conditions. SYBR® Green I, on the other hand, is known to degrade following multiple freeze-thaw cycles and under PCR conditions. Moreover, decomposed SYBR® Green I is reported to be even more inhibitory to PCR.
Digital PCR: EvaGreen® dye is currently the only qPCR dye to be used in droplet digital PCR (ddPCR).
For most qPCR applications, we recommend using EvaGreen® Dye, 20X in water (31000), or one of our 2X EvaGreen® qPCR Master Mixes. EvaGreen® Dye, 2000X in DMSO (31019) is offered for specialized applications requiring a concentrated dye solution.
N-Flux™ 5X Digital PCR Master Mix is a high-performance, dye-free master mix specially formulated to address the complexities of reagent partitioning within microfluidic digital PCR plates and chips. Developed for chip-based digital PCR (cdPCR) instruments in collaboration with Combinati, we offer this master mix for digital PCR assays and R&D development of novel dPCR and qPCR platforms.
Sensitive, robust, and reproducible performance in dPCR and qPCR assays
Minimizes interactions with hydrophobic plastic surfaces for optimal sample partitioning
Maximizes end-point fluorescence for superior separation between positive and negative partitions
Validated for multiplex or singleplex, genotyping, and copy number variation dPCR and qPCR
Suitable for all fluorescent probe types, including TaqMan®, BHQ®, and molecular beacons
5X concentration facilitates PCR with dilute or large sample volumes
Formulated with Cheetah™ HotStart Taq for fast cycling protocols
N-Flux™ 5X Digital PCR Master Mix
ROX Reference Dye
A Novel Master Mix Optimized for Microfluidics
N-Flux™ 5X Digital PCR Master Mix is designed to optimize partitioning and minimize sample adsorption during flow through the microfluidic channels of the chips and plates used in cdPCR. The master mix maximizes end-point fluorescence for superior separation between positive and negative partitions resulting in sensitive and accurate results. N-Flux™ 5X Digital PCR Master Mix has been validated in singleplex and multiplex probe-dPCR reactions. Using the Combinati Absolute Q® digital PCR platform, N-Flux™ 5X Digital PCR Master Mix reproducibly performed genotyping and copy number variation analysis with quantitative discrimination. The master mix is suitable for all fluorescent probe-based technologies, including hydrolysis probes (such as TaqMan® and dual-labeled BHQ® probes) and displacement probes (like molecular beacons). Contact usto inquire about custom formulations for your platform development project.
While N-Flux™ 5X Digital PCR Master Mix is formulated to address the complexities of flow through microfluidic chambers, the 5X concentration can also simplify the work flow for qPCR users. The high concentration gives flexibility and facilitates PCR analysis of large sample volumes or samples that are too dilute for traditional 2X master mixes. See our Forget-Me-Not™ Master Mixes for EvaGreen® dye-based or probe-based quantitative real-time PCR.
The kit contains master mix with Cheetah™ HotStart Taq DNA Polymerase and dNTPs in a specially formulated buffer and includes a separate vial of ROX passive reference dye for optional use.
Cheetah™ HotStart Taq DNA Polymerase
Biotium’s proprietary chemically modified hot-start DNA Polymerase. Cheetah™ Taq is completely inactive at room temperature, but is fully activated in 2 minutes at 95°C with high activity recovery, making it particularly suitable for fast PCR.
Absolute Q is a registered trademark of Combinati; BHQ is a registered trademarks of LGC Biosearch Technologies; TaqMan is a registered trademark of Roche Diagnostics.