Forget-Me-Not qPCR Master Mix with ROX



Product Description

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

  • <40Ȼ 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 <40Ȼ -55Ȼ 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.

Left: 1X Forget-Me-Not™ Master Mix. Right: 1X Forget-Me-Not™ Master Mix after addition of Forget-Me-Not™ Template Buffer.
Left: 1X Forget-Me-Not™ Master Mix. Right: 1X 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 with ROX (31042) includes a 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. We also offer Forget-Me-Not™ qPCR Master Mix (31041) without ROX for use with instruments that do not require ROX reference dye.


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 €œrelease-on-demand€ mechanism, which permits the use of a relatively high dye concentration in qPCR without PCR inhibition (Ref. 1). 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 (Ref. 2). 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 qPCR
Figure 2. Real-time PCR data comparing Forget-Me-Not™ (blue lines) with Biotium’s Fast EvaGreen® (green lines) and Qiagens QuantiNova® SYBR® Green (gray lines) master mixes. A. Amplification curves on linear scale. EvaGreen® dye-based master mixes yield higher signal compared to the SYBR® Green-based mix. B. Amplification curves on logarithmic scale. Forget-Me-Not™ performs as well or better than the other master mixes. NTCs: non-template controls (dashed lines).

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 other qPCR  reagents and kits.

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.

1. Mao, et al. Characterization of EvaGreen Dye and the implication of its physicochemical properties for qPCR applications. BMC Biotechnology 7, 76-91 (2007).
2. Ohta, et al. Ethidium bromide and SYBR Green I enhance the genotoxicity of UV-irradiation and chemical mutagens in E. coli. Mutation Res. 492, 91-97 (2001).

MSDS (PDF): MSDS 31041, 31042
Protocol (PDF): PI-31041, 31042
Safety Report (PDF): EvaGreen Safety Report

Shipment Method: Shipping and handling methods will be assessed and calculated at time of shipment based upon item(s) storage temperature conditions.

Expedited shipment may be requested at time of checkout.

Please note that products will be shipped at ambient temperature. This will not affect performance if the products are kept at the indicated temperature for long term storage. Please refer to the package insert for correct storage conditions.

More information about product handling and shelf life