描述For qPCR users using SYBR green dye who need to minimize optimization time, Applied Biosystems PowerUp SYBR Green Master Mix offers proprietary Dual-Lock hot start technology. Unlike other polymerases, a Dual-Lock polymerase offers two unique hot start mechanisms to prevent primer dimer and non-specific amplification.PowerUp SYBR Green Master Mix is a pre-formulated, optimized, universal 2X master mix for real-time PCR workflows. Coupled with user-supplied primer sets and template, PowerUp SYBR Green Master Mix is designed to amplifiy targets for accurate gene expression analysis. PowerUp SYBR Green Master Mix features include: • A dual hot-start mechanism for excellent specificity • Highly reproducible CTs over a broad dynamic range • Inclusion of UDG to help prevent carryover contamination • Stability of pre-assembled reactions for up to 72 hours • Compatibility with most real-time qPCR instruments Exceptional specificity with dual hot-start mechanism Specificity is paramount in obtaining high-quality data in SYBR Green reactions and can be enhanced through control of the Taq DNA polymerase at lower temperatures before stringent primer binding. PowerUp SYBR Green Master Mix uses the Dual-Lock Taq DNA polymerase enzyme that combines two unique hot-start mechanisms to help control its activity and to prevent undesirable early activity of the polymerase at low temperatures. In an experiment evaluating 24 different primer sets, PowerUp SYBR Green Master Mix generated single melt curves 100% of the time, consistent with highly specific amplification. Tight reproducibility in CTs over a wide dynamic range PowerUp SYBR Green Master Mix demonstrates excellent reproducibility over a wide dynamic range for a variety of targets tested (see figure). In addition, this master mix provides efficient amplification over 6 logs of sample input. Unlike competitor mixes that can exhibit inhibition at high cDNA inputs and low correlation coefficients over a dynamic range experiment, PowerUp SYBR Green Master Mix efficiency is maintained over the full dynamic range to help deliver maximal confidence in data quality (see figure). UDG included for carryover contamination control1 The inclusion of uracil-DNA glycosylase (UDG) and dUTP in the PowerUp SYBR Green Master Mix enables the degradation of any previously amplified PCR products, helping prevent contamination of subsequent qPCR reactions and possible false positives. Stability of pre-assembled reactions The tight control of the Dual-Lock Taq DNA polymerase in PowerUp SYBR Green Master Mix enables reactions to be assembled up to 72 hours prior to cycling without impacting data (see figure). This consistency allows users to have confidence and flexibility in the use of PowerUP SYBR Green Master Mix across numerous workflow scenarios.2 Broad instrument compatibility PowerUp SYBR Green Master Mix can be used in either standard or fast cycling mode and is compatible with all Applied Biosystems real-time PCR instruments.3 It is also compatible with the Bio-Rad IQ™5 and CFX96™/CFX384™ systems, Roche LightCycler™ LC480, and Stratagene™ MX3005P™ systems. For optimal results, recommended primer concentrations are 300–800 nM. Learn more about PowerUp SYBR Green Master Mix > 1 Master mixes containing UDG should not be used with bisulfite-treated DNA samples. 2 Pre-PCR stability is influenced not only by the master mix, but also by the target being analyzed. For maximum confidence, researchers should confirm stability profiles of their specific targets. 3 For optimal performance, standard cycling using the Applied Biosystems 7900 system is recommended. |
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