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  7. Applied Biosystems™ PowerUp™ SYBR™ Green Master Mix for qPCR

Applied Biosystems™ PowerUp™ SYBR™ Green Master Mix for qPCR

Catalog No. A25741
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Catalog No. Quantity
A25741 1 mL
A25742 5 mL
A25743 50 mL
A25778 10 x 5 mL
A25780 5 x 1 mL
A25918 10 x 1 mL
A25776 2 x 5 mL
A25777 5 x 5 mL
A25779 2 x 1 mL
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Catalog No. A25741 Supplier Applied Biosystems™ Supplier No. A25741
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Description

Pre-formulated, optimized, universal 2X master mix for real-time PCR workflows.

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 control

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 scenarios¹.

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 instruments². 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.

¹Pre-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.

² For optimal performance, standard cycling using the Applied Biosystems™ 7900 system is recommended.

Order Info

Shipping Conditions: Wet Ice

Specifications

Concentration 2X
Content And Storage The 2X mix contains SYBR Green dye, Dual-Lock Taq DNA Polymerase, dNTPs with dUTP/dTTP blend, heat-labile UDG, ROX passive reference dye, and optimized buffer components. Contains 1 x 1 mL tube, sufficient for 100 20-μL reactions (10 μL Master Mix per reaction).

Store at 2–8°C in the dark.
Detection Method SYBR
Format Tube
PCR Method qPCR
Polymerase Dual-Lock Taq DNA Polymerase
Reaction Speed Fast or Standard
For Use With (Equipment) 7500 Fast System, 7500 System, 7900HT System, QuantStudio 12k Flex, QuantStudio 3, QuantStudio 5, QuantStudio 6 Flex, QuantStudio 6 Pro, QuantStudio 7 Pro, QuantStudio 7 Flex, StepOne, StepOnePlus, ViiA 7 System
Multiplex Capability Single-plex
Passive Reference Dye ROX (Pre-mixed)
Product Line PowerUP, SYBR
Product Type Real Time PCR SYBR Master Mix
Quantity 1 mL
Shipping Condition Wet Ice
Sufficient For 100 Reactions of 20 μL
Target Specificity cDNA or gDNA only
For Use With (Application) Gene Expression
Thermostability 72 hr., Assembled Reaction
No. of Reactions 100 Reactions
Sample Type DNA
Volume 1 mL
Includes dUTP, UNG/UDG
Inhibitor Tolerance (Simple) Medium
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Frequently Asked Questions (FAQs)

When using the PowerUp SYBR Green Master Mix (Cat. No. A25741, A25742, A25743, A25776, A25777, A25778, A25779, A25780, A25919), I get high Ct values. What can I do?

It could be due to the following reasons:

  • Insufficient cDNA template is present. Use 10 to 100 ng of cDNA template per 50 µL reaction.
  • Quality of cDNA template is poor. Quantify the amount of cDNA template and test the cDNA template for the presence of PCR inhibitors.
  • Sample degradation. Prepare fresh cDNA, then repeat the experiment.
  • Reduced number of PCR cycles in the thermal cycling protocol. Increase the number of PCR cycles to the default setting of 40.
  • Primer-dimer formation and residual polymerase activity. Prepare the reaction mixes and reaction plate on ice. To ensure optimal results, run the reaction plate as soon as possible after completing the reaction setup.
When using the PowerUp SYBR Green Master Mix (Cat. No. A25741, A25742, A25743, A25776, A25777, A25778, A25779, A25780, A25919), the Multicomponent signal for ROX is not linear. What could cause this?

This could be due to the following reasons:

  • Pure dye component’s spectra are incorrect. Rerun pure dye spectra.
  • Incorrect dye components have been chosen. Choose correct dyes for data analysis.
Which reverse transcription kit can I use with the PowerUp SYBR Green Master Mix (Cat. No. A25741, A25742, A25743, A25776, A25777, A25778, A25779, A25780, A25919)?

We recommend using the High-Capacity RNA-to-cDNA Kit (Cat. No. 4387406, 4388950).
What primer concentrations can I use with the PowerUp SYBR Green Master Mix (Cat. No. A25741, A25742, A25743, A25776, A25777, A25778, A25779, A25780, A25919)?

The recommended primer concentration range is 300-800 nM. You will need to optimize the primer concentration by running different concentration combinations of the forward and reverse primer. See page 28 of the PowerUp SYBR Green Master Mix User Guide.
Does the PowerUp SYBR Green Master Mix (Cat. No. A25741, A25742, A25743, A25776, A25777, A25778, A25779, A25780, A25919) contain uracil-DNA glycolase?

This master mix has a heat-labile UDG. It is completely and irreversibly inactivated after 10 minute incubation at 50°C.
Which enzyme does the PowerUp SYBR Green Master Mix (Cat. No. A25741, A25742, A25743, A25776, A25777, A25778, A25779, A25780, A25918) have? What is the advantage of this enzyme?

The PowerUp SYBR Green Master Mix uses a Dual Lock Taq DNA Polymerase enzyme. This enzyme combines two unique hot-start mechanisms to help control its activity and to prevent undesirable early activity of the polymerase at low temperatures. As such, this mechanism provides excellent specificity.
Can I store PowerUp SYBR Green Master Mix for qPCR at -20 degrees C?

We have not tested PowerUp SYBR Green Master Mix for qPCR at -20 degrees C. We have only performed accelerated and real-time stability testing at 2-8 degrees C, as this is the recommended and most convenient storage temperature.

What is the recommended storage temperature for the PowerUp SYBR Green Master Mix?

The recommended storage temperature for the PowerUp SYBR Green Master Mix is 2-8 degrees C.
What is the difference between Power SYBR Green Master Mix and PowerUp SYBR Green Master Mix?

Both master mixes are compatible with all Applied Biosystems Real Time PCR instruments, but Power SYBR Green Master Mix can only be run in Standard Mode. PowerUp SYBR Green Master Mix contains a heat-labile UDG, which allows for review of PCR products up to 72 hrs post-PCR (unlike Power SYBR Green Master Mix). Finally, PowerUp SYBR Green Master Mix uses Dual-Lock Taq DNA Polymerase, which uses a combination of two hot start modifications for exceptional specificity.
Which instruments are compatible with the PowerUp SYBR Green Master Mix?

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 (except the 7900HT Fast mode instrument). It is also compatible with the Bio-Rad iQ5, Roche LightCycler 480, and Agilent MX3005P systems.

For Research Use Only. Not for use in diagnostic procedures.