Forget-Me-Not™ EvaGreen® qPCR Master Mix
Forget-Me-Not™ EvaGreen® master mix features:
- As low as $20/mL for the 20 mL 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™ 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 can also be used without the Template Buffer if you prefer.
Forget-Me-Not™ EvaGreen® Kit Components:
2X Forget-Me-Not™ EvaGreen® 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™ EvaGreen® qPCR Master Mix comes in two options: Cat. # 31041 does not include ROX reference dye, and for instruments that require either a low or high concentration of ROX reference dye, Cat. # 31042 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™ EvaGreen® qPCR Master Mix Protocol for details.
Forget-Me-Not™ Universal Probe Master Mix
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.
Forget-Me-Not™ Universal Probe Kit Components:
2X Forget-Me-Not™ Universal Probe Master Mix
The 2X Master Mix contains Cheetah™ HotStart Taq DNA Polymerase and dNTPs. The user provides the probe of their choice.
Forget-Me-Not™ Tracking Buffer
The 40X Forget-Me-Not™ Tracking Buffer contains an inert blue dye. You have the choice of adding Tacking 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. On the other hand, adding Tracking Buffer to your DNA template samples allows you to track which reactions have had template added while you set up your PCR reactions.
ROX Reference Dye
Forget-Me-Not™ Universal Probe Master Mix comes in two options: Cat. # 31043 does not include ROX reference dye, and for instruments that require either a low or high concentration of ROX reference dye, Cat. # 31044 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™ Universal Probe Master Mix Protocol for details.
Unique Forget-Me-Not™ Features:
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.
EvaGreen® Dye and Cheetah™ HotStart Taq DNA Polymerase are covered under US and international patents. QuantiNova is a registered trademark of Qiagen Group. SYBR 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.