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5ml
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.
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文献和实验SYBR Green Quantitative PCR Protocol
12μl of master mix for each well, plus some excess1 Rxn: 10μl SYBR Green Mix 52 Rxn: 520μl SYBR 0.4μl Forward Primer (10μM stock) 20.8μl F
REAL TIME PCR WITH SYBR GREEN I 建议PROTOCOL
1、反应体系 25ulCocktail 20 ul: Primer FP 1 ul + Primer RP 1u + 2 * SYBR GREEN I MIX 12.5 + H2O 5.5 ul cDNA 5 ul2、Primer 浓度，需要摸索，从200nM到700nm等，设置不同浓度。3、每个引物浓度，从well 1到12设置4个样本，阳性cDNA 1和2，NTC以及H2O，做triplicate。4、若用ABI7000或7700，所有的PCR条件可设置成一致的，减少麻烦，当然，引物
Vybrant® DyeCycle Green and Orange Stains
Stability Vybrant® DyeCycle™ Green stain Vybrant® DyeCycle™ Orange stain 400 μL 5 mM solution in dimethyl sulfoxide (DMSO
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