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.
The master mix contains Cheetah™ HotStart Taq DNA Polymerase
and dNTPs in a buffer optimized for high qPCR sensitivity and multiplex reactions. Only primers, probe and template need to be added.
Cheetah™ HotStart Taq DNA Polymerase is Biotium’s proprietary chemically modified hot-start DNA Polymerase. Cheetah™ Taq is fully activated in 2 minutes with high activity recovery, making it particularly suitable for fast PCR. Cheetah™ Taq is completely inactive at room temperature.
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.
SNP genotyping using dual-labeled BHQ1Plus® hydrolysis probes. DNA samples were genotyped in reactions containing 450 nM of each primer and 100 nM of each probe (Allele 1 probe-FAM/ BHQ®-1, Allele 2 probe- CAL Fluor® Orange 560/BHQ®-1) on a QuantStudio® 5 system (ThermoFisher Scientific).