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.
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.
Note: 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 (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.
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).
Biotium公司是一家在荧光技术领域独树一帜的生物技术公司,公司位于美国加利福尼亚的BAY AREA------世界上首要的生物科技中心。该公司一直致力于开发和生产用于生物医学研究的荧光指示剂及其他特殊生化产品。其产品被广泛应用于细胞生物学、蛋白质组学、分子生物学、免疫学、微生物学、诊断学、生物化学、神经科学、血液流动检测及大规模药物筛选等众多领域。Biotium公司专利产品众多,包括已经被广泛使用的实时PCR定量荧光染料EvaGreen、具有划时代意义的凝胶核酸染料GelRed和GelGreen,以及最新推出的Cheetah Taq热启动酶个CF系列蛋白质标记染料等等。