Forget-Me-Not™ EvaGreen® qPCR Master Mix (2-Color Tracking)

2X Forget-Me-Not™ EvaGreen® qPCR Master Mix is a ready-to-use high performance dye-based master mix for qPCR and DNA melt curve analysis. 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 easily distinguish wells containing reaction mix from empty wells. For 2-color tracking, mix your template with Forget-Me-Not™ Template Buffer to see at a glance which wells have master mix or template.

Note: We also offer 2X Master Mix (99801-10ML) for standalone purchase for customers who do not wish to use template buffer.

• Ready-to-use, just add primers and DNA template
• Superior EvaGreen® Dye is more sensitive than SYBR® Green I and is compatible with HRM®
• Prevent wasted samples and costly mistakes with Forget-Me-Not™ 2-color tracking
• Formulated with Cheetah™ HotStart Taq for fast cycling protocols
• Directly visualize amplification product fluorescence in a gel
• No ROX or Separate ROX for use with any PCR instrument

Figure 1. Left: 1X Forget-Me-Not™ Master Mix. Right: 1X Forget-Me-Not™ Master Mix after addition of DNA template mixed with Forget-Me-Not™ Template Buffer.

Prevent Waste & Catch Mistakes

Catch pipetting mistakes and avoid wasting time, reagents, and your precious DNA samples. The 2-color tracking Forget-Me-Not™ EvaGreen® qPCR Master Mixes features a unique combination of a master mix containing a low concentration of blue dye and a DNA template buffer containing a high 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).  The Forget-Me-Not™ Template Buffer is optional if you do not require 2-color tracking. You can also use Forget-Me-Not™ EvaGreen® qPCR Master Mix (Low ROX or High ROX), which are premixed with ROX and allow single-color tracking.

Peace of Mind Plus PCR Performance

Forget-Me-Not™ qPCR Master Mix is formulated for fast cycling PCR parameters, but can also be used with regular cycling protocols. It performs as well as our Fast EvaGreen® Master Mix, and as well 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.

Figure 2. Real-time PCR data comparing Forget-Me-Not™ (blue lines) with Biotium’s Fast EvaGreen® (green lines) and Qiagen’s 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).

EvaGreen® Dye

An environmentally friendly, non-mutagenic, and non-cytotoxic DNA-binding dye with features ideal for both qPCR and High Resolution Melt® (HRM) analysis. EvaGreen® Dye binds to dsDNA via a novel “release-on-demand” mechanism, which prevents dye transfer from amplicon to amplicon and permits the use of a relatively high dye concentration in qPCR without PCR inhibition. Learn more about EvaGreen® Dye for qPCR.

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.

ROX Reference Dye

ROX is required as a passive reference dye in some qPCR systems to compensate for well-to-well optical variations. Refer to the table below for the passive dye requirements of real-time PCR instruments. Product #31041 is supplied without ROX, while product #31042 is supplied with a separate vial of ROX.  For master mixes with pre-mixed ROX, use Forget-Me-Not™ EvaGreen® qPCR Master Mix (Low ROX or High ROX)