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Forget-Me-Not™ Universal Probe qPCR Master Mix

A high-performance product for fluorescent probe-based PCR applications, including quantitation and SNP genotyping.

Reference dye
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1 mL (100 reactions) Trial Size
5 x 1 mL (500 reactions)
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Product Description

2X Forget-Me-Not™ Universal qPCR Probe Master Mix is a high-performance product for fluorescent probe-based PCR applications, including quantitation and SNP genotyping.

  • Suitable for all fluorescent probe types, including TaqMan®, BHQ®, and molecular beacons
  • Sensitive, robust, and reproducible performance at a fraction of the cost of other mixes
  • For singleplex or multiplex reactions
  • Tracking Buffer helps you catch pipetting mistakes, saving time and precious samples
  • Cheetah™ HotStart Taq for fast cycling protocols
  • With or without separate ROX for use with any qPCR instrument

The master mix contains Cheetah™ HotStart Taq DNA Polymerase and dNTPs in a buffer optimized for high qPCR sensitivity and multiplex reactions. Only primers, probe, and template need to be added.

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.

Forget-Me-Not™ Tracking Buffer

The 40X Forget-Me-Not™ Tracking Buffer contains an inert blue dye. You have the choice of adding Tracking Buffer to the master mix, to the DNA template, or not to use the tracking buffer in your reactions. Addition of Tracking Buffer to your master mix allows you to easily distinguish wells containing reaction mix from empty wells. Alternatively, when added to your DNA template samples, the Tracking Buffer allows you to see which reactions have had template added while you set up your PCR reactions.

SNP genotyping using dual-labeled BHQ1Plus® hydrolysis probes. DNA samples were genotyped in reactions containing 450 nM of each primer and 100 nM of each probe (Allele 1 probe-FAM/ BHQ®-1, Allele 2 probe- CAL Fluor® Orange 560/BHQ®-1) on a QuantStudio® 5 system (ThermoFisher Scientific).

Singleplex and triplex PCR of GAPDH, ACTB, and TFCR from human genomic DNA. PCR detection of GAPDH, ACTB, or TFCR in singleplex reactions, or together in triplex reactions. Reactions contained 20 ng of human genomic DNA, 400 nM of each primer, and 200 nM of each hydrolysis probe (GAPDH-FAM/BHQ®-1, ACTB-CAL Fluor® Orange 560/BHQ®-1, TFCR-CAL Fluor Red 610/ BHQ®-2.

 

Cheetah Taq is covered under US and international patents. TaqMan is a registered trademark of Roche Diagnostics. CAL Fluor and BHQ are registered trademarks of LGC Biosearch Technologies.

Product Attributes

Reference dye
No ROX, With Separate ROX
Detection method/readout
PCR/qPCR
Size
1 mL (100 reactions) Trial Size, 5 x 1 mL (500 reactions)

Documents, Protocols, SDS and COA

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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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