Dye based One Step RT-qPCR with UNG & MegaMix Emerald

Containing all the components needed to perform RT-qPCR swiftly and reliably.

The kit consists of 2X MME Mastermix, containing chemically modified Hot Start Taq DNA polymerase, intercalating dye, dUTP and UNG in enhancing buffer optimised; 20X RTase/RI enzyme mix, optimised for amplifying low copy RNA targets; and Just Water, our molecular biology grade water. The third generation intercalating fluorescent dye binds to double stranded DNA, making MegaMix Emerald the perfect choice for qPCR, Melt Curve Analysis and High-Resolution Melting (HRM). The Hot Start Taq polymerase is chemically inactivated until heating to 95°C, providing excellent sensitivity and specificity, eliminating the formation of non-specific amplification and primer-dimers. The presence of UNG and dUTP’s eliminates carryover contamination, the thermolabile UNG is active at room temperature before being completely and irreversibly inactivated when heated to 95°C, this means your PCR product is suitable for downstream processing.

Key Features

• Specificity—Hot Start Taq DNA Polymerase in optimized buffer eliminates non-specific amplification and the formation of primer dimers.
• Sensitivity—Single copy detection.
• Eliminates carry over contamination—incorporation of thermolabile UNG and dUTP prevent amplicon contamination from previous runs without degrading generated amplicons.
• Versatile— compatible with standard and fast cycling conditions, GC/AT rich templates.
• Reproducibility and convenience—ready to use 2X format.
• Inhibitor Resistance—suitable for direct to PCR with products such as microLYSIS-RNA.
• Third generation intercalating dye— no inhibition of PCR, even at high concentrations, excellent for High Resolution Melting.

Applications

• Quantification of any RNA template (mRNA, total RNA, viral RNA), low copy number samples.
• Gene expression analysis.
• Pathogen detection.
• RT-qPCR assays using fluorescence of intercalating dye.
• High resolution melting.