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N-Flux™ 5X Digital PCR Master Mix

A 5X master mix specially formulated to address the complexities of reagent flow through dPCR chips and plates to achieve optimal sample partitioning and maximum end-point fluorescence.

 

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Trial size (1 x 200uL)
5 x 200uL
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Product Description

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

Kit Components

  • 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 us to inquire about custom formulations for your platform development project.

High-resolution, multiplex copy number variation analysis performed using N-Flux™ 5X Digital PCR Master Mix on the Combinati Microfluidic Array Partitioning (MAP16) device. Superior partitioning results in quantitative determination of the copy number of a low abundance SNP variant and a high abundance control on human gDNA, with excellent separation between positive and negative wells.

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.

Product Attributes

Size
Trial size (1 x 200uL), 5 x 200uL
Reference dye
With Separate ROX
Storage Conditions
Store at -10 to -35 °C, Protect dye component from light
Shelf life
Guaranteed for at least 12 months from date of receipt when stored as recommended

Documents, Protocols, SDS and COA

FAQs

EvaGreen® Dye and Master Mixes

The ROX concentration is 35 nM which is suitable for low ROX instruments. If you are using a high ROX instrument, you can add ROX, 25 mM in TE Buffer (Catalog # 29052) which is available separately (Cat # 29052).

Our original EvaGreen® Dye is a saturating dsDNA binding dye that is superior for quantitative real-time PCR (qPCR), high-resolution melt (HRM), digital droplet PCR (ddPCR) and other genomics applications over SYBR® Green I and other commercial PCR dyes. It offers several essential features critical for PCR and related applications including high thermal, chemical and photostability, high sensitivity due to high signal to noise related to its novel ‘release-on-demand’ mechanism, non-inhibitory to PCR, and lack of dye migration.  In addition, EvaGreen® Dye is non-toxic, non-mutagenic, and not hazardous to aquatic life.

EvaGreen® Plus Dye is an advanced version of the original EvaGreen® Dye, retaining the same essential benefits while providing a higher signal-to-noise ratio. This greater sensitivity offers further advantages for digital PCR and isothermal applications.

EvaGreen® Dye is used in all of Biotium’s dye-based qPCR master mixes, which combine superior brightness and sensitivity, with the ability to do melt curve analysis in the same reaction. The new EvaGreen® Plus Dye is available as a stand-alone 20X solution in water.

0.5X to 1X EvaGreen® is commonly used for LAMP applications when fluorescent signal is read by instrumentation. For detection by eye, an increased concentration of EvaGreen® can be used. For example,  Lee, S. H. et al. tested up to 100X EvaGreen® and found that 10X EvaGreen® was optimal for their assay.

Lee, S. H. et al. Front. Microbiol., 09 January 2017 | https://doi.org/10.3389/fmicb.2016.02166

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