Quick links
Frequently asked questions - categories
Ask a question
Contact Us

Forget-Me-Not™ qPCR Master Mixes


High performance master mix with 2-color reaction component tracking system, at a great price

  • <40⊄/reaction when you buy the 500 reaction size
  • 2-color system helps you catch pipetting mistakes, saving time, reagents, DNA samples, and $$$
  • EvaGreen® Dye – better signal than SYBR® Green, safer and more environmentally friendly
  • Cheetah™ Hotstart Taq – fully activated after two minutes at 95°C for fast cycling protocols

Forget-Me-Not™ is a high performance qPCR master mix available for a fraction of the cost of similar products, regularly priced at <40⊄-55⊄/reaction, vs $0.50-$1.00/reaction for color tracking mixes from other suppliers.

Forget-Me-Not™ qPCR Master Mix is a hot-start EvaGreen® dye-based master mix for use in real time PCR applications and DNA melt curve analysis. Forget-Me-Not™ features a unique combination of a master mix containing a low concentration of blue dye, plus a DNA template buffer containing a higher concentration of blue dye. When you add the 2X Forget-Me-Not™ Master Mix to your reaction, it appears light blue (Figure 1, left). Then, when you add template containing Forget-Me-Not™  Template Buffer to the reaction, the color turns dark blue (Figure 1, right). Forget-Me-Not™ allows you to see at a glance whether you forgot to add master mix or template to any of your reactions, so you can catch pipetting mistakes and avoid wasting time, reagents, and your precious DNA samples. The 2X Forget-Me-Not™ Master Mix also can be used without the Template Buffer if you prefer.

Figure 1. Forget-Me-Not™ allows you to see at a glance whether you’ve added reaction components to your tubes or wells. Left: 1X Forget-Me-Not™ Master Mix; Right: Forget-Me-Not™ Master Mix after addition of Forget-Me-Not™ Template Buffer.

Kit Components:

Forget-Me-Not™ qPCR Master Mix
The 2X Master Mix contains EvaGreen® dye, Cheetah™ HotStart Taq DNA Polymerase, dNTPs, and a low concentration of an inert blue dye, which allows the user to distinguish wells containing reaction mix from empty wells. You can use the master mix with or without the Forget-Me-Not Template Buffer.

Forget-Me-Not™ Template Buffer
The 40X Template Buffer contains a high concentration of inert blue dye. Adding the template buffer to your DNA template samples allows you to easily track which reactions have had template added when your are setting up your PCR reactions.

ROX Reference Dye
Forget-Me-Not™ qPCR Master Mix (31041) does not include ROX reference dye. For instruments that require either a  low or high concentration of ROX reference dye, we offer Forget-Me-Not™ qPCR Master Mix with ROX (31042), which includes a separate vial of ROX Reference dye, and instructions for how much ROX to add depending on which protocol and instrument you are using. Download the Forget-Me-Not™ qPCR Master Mix Protocol for details.


EvaGreen® dye

A unique DNA-binding dye with features ideal for both qPCR and High Resolution Melting® (HRM) analysis*. EvaGreen® dye binds to dsDNA via a novel “release-on-demand” mechanism, which permits the use of a relatively high dye concentration in qPCR without PCR inhibition (Mao, et al. 2007. BMC Biotech. 7, 76-91).

A unique feature of EvaGreen® dye is its safety. DNA-binding dyes are inherently dangerous due to their potential to cause mutation. With this in mind, Biotium’s scientists designed EvaGreen® dye such that it cannot cross cell membranes, thus preventing the dye from being in contact with genomic DNA in live cells. All other commercial PCR dyes enter into cells in a matter of minutes. SYBR® Green I, for example, has been shown to be environmentally more toxic than ethidium bromide, a well-known mutagen (Ohta, et al. 2001. Mut. Res. 492, 91-97). Independent labs have confirmed that EvaGreen dye is nonmutagenic, noncytotoxic and safe to aquatic life for direct disposal in the drain. For details, download the EvaGreen® Dye Safety Report.

An added benefit of an EvaGreen®-based master mix is that you can analyze your PCR product by gel electrophoresis without the need to add another DNA-binding dye to either your loading buffer or gel. The EvaGreen® dye in the master mix can act as a DNA pre-stain, permitting direct visualization of DNA bands following electrophoresis.

Cheetah™ HotStart Taq

Cheetah™ HotStart Taq DNA Polymerase is Biotium’s proprietary chemically-modified hot-start DNA Polymerase. Cheetah™ Taq is completely inactive at room temperature, and is fully activated after 2 minutes at 95°C, making it particularly suitable for fast cycling PCR protocols.

FMN, Fast EG, QuanitNova comparison
Figure 2. Comparison of GAPDH fast-cycling real time qPCR assay using Forget-Me-Not™ qPCR Master Mix, Biotium’s Fast EvaGreen® qPCR Master Mix, or Qiagen’s QuantiNova® SYBR® Green qPCR Master Mix. A. Raw fluorescence data. EvaGreen®-based master mixes yield higher fluorescence signal compared to the SYBR® Green-based master mix. B. Data plotted on logarithmic scale. Forget-Me-Not™ performs as well or better than other PCR master mixes.

Peace of mind, plus PCR performance

Forget-Me-Not™ qPCR Master Mix has been formulated for fast cycling PCR parameters, but can also be used with regular cycling protocols. Forget-Me-Not™ qPCR Master Mix performs as well as our Fast EvaGreen® Master Mix, and as well as or better than Qiagen’s QuantiNova® SYBR® Green PCR Master Mix in a real-time PCR assay (Figure 2), while making  it easy for you to make sure your reactions are set up correctly the first time, every time.

Also see Biotium’s Fast EvaGreen®, Fast Plus EvaGreen®, and Fast Probe qPCR Master Mixes.

Forget-Me-Not™ Universal Probe Master Mix

2X Forget-Me-Not™ Universal Probe Master Mix is a high-performance product for fluorescent probe-based PCR applications, including quantitation and SNP genotyping. This kit 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). Forget-Me-Not™ Universal Probe Master Mix shows excellent concordance of results in singleplex and multiplex reactions, has broad instrument compatibility, and can be used in both standard and fast protocols. A separate kit is also available with ROX, if your instrument requires ROX reference dye.

Product NameCatalog NumberFeatures
Forget-Me-Not qPCR Master Mix31041EvaGreen dye-based master mix with a unique two-color system for tracking which reagents you've added to your reaction.
Forget-Me-Not qPCR Master Mix with ROX31042EvaGreen dye-based master mix with a unique two-color system for tracking which reagents you've added to your reaction, with ROX reference dye.
Forget-Me-Not Universal Probe Master Mix31043High-performance master mix for fluorescent probe-based PCR applications, including quantitation and SNP genotyping.
Forget-Me-Not Universal Probe Master Mix with ROX31044High-performance master mix for fluorescent probe-based PCR applications, including quantitation and SNP genotyping. With ROX reference dye.


EvaGreen® Dye and Cheetah™ HotStart Taq DNA Polymerase are covered under US and international patents. QuantiNova® is a registered trademark of Qiagen Group. SYBR® Green is a registered trademark of Molecular Probes, Inc./Life Technologies. *HRM® is a registered trademark of Idaho Technologies, Inc./BioFire Defense, LLC and its use may require a license.