PerfeCTa SYBR Green SuperMix, UNG

PerfeCTa® SYBR® Green SuperMix, UNG is a 2X concentrated, ready-to-use reaction cocktail that contains all components, except primers and template for real-time quantitative PCR systems that do not require an internal reference dye. The proprietary buffer and stabilizers have been optimized exclusively for SYBR® Green I qPCR to deliver maximum PCR efficiency, sensitivity, and robust fluorescent signal. Highly specific amplification is crucial to successful qPCR with SYBR® Green I technology because this dye binds to and detects any dsDNA generated during amplification. A key component of this supermix is AccuStart™ Taq DNA polymerase which contains monoclonal antibodies that bind to the polymerase and keep it inactive prior to the initial PCR denaturation step. Upon heat activation (2 minutes at 95°C), the antibodies denature irreversibly, releasing fully active, unmodified Taq DNA polymerase. This enables specific and efficient primer extension with the convenience of room temperature reaction assembly. Additionally, the dNTP mix in this SuperMix contains dUTP in place of dTTP. Inclusion of uracil-N-glycosylase (UNG) prevents amplification of carry-over contamination from previous dU-containing PCRs.

• 2X concentrated master mixes with stabilized SYBR Green dye and exceptional temperature stability (≥30 days at 22°C) that withstand repetitive freeze-thaw (≥ 20X).
• Maximum SYBR Green dye load for robust optical signal with small amplicons. · Supports efficient vortexmixing with proprietary anti-foaming technology.
• Maximum assay sensitivity and target precision with highly modified Taq DNA polymerase and stringentultrapure, AccuStart™ II antibody hotstart technology.
• Easy-to-use with AccuVue™ plate loading dye and pre-blended passive reference dye.

Features & Benefits

• Buffer optimized for SYBR® Green stability, ensures reliable performance
• High yield, high sensitivity
• Highly specific amplification
• Superior reproducibility and precision

Performance Data

Comparison to QuantiTect, GAPD

PerfeCTa SYBR Green SuperMix Comparison to QuantiTect SYBR Green PCR Kit (Qiagen), GAPD
Log-fold serial dilutions of qScript™ cDNA from HeLa cell total RNA (100 ng to 0.1 pg) were analyzed by SYBR Green qPCR following each manufacturers protocol. Averaged plots of triplicate qPCR reactions are shown.

Real-Time PCR Instrument Compatibility

• Roche LightCycler™ 480
• Qiagen/Corbett Rotor-Gene® Q
• Qiagen/Corbett Rotor-Gene® 6000
• Qiagen/Corbett Rotor-Gene® 3000
• Eppendorf Mastercycler® ep realplex
• Eppendorf Mastercycler® ep realplex2 s
• Cepheid SmartCycler®
• Bio-Rad CFX96™
• Bio-Rad CFX384™
• Bio-Rad/MJ Opticon®
• Bio-Rad/MJ Opticon 2
• Bio-Rad/MJ Chromo4™
• Bio-Rad MiniOpticon™
• Illumina Eco qPCR
• Thermo Scientific PikoReal Cycler

Contents

• Single-tube, 2X concentrated reagent containing:
  • Reaction buffer with optimized concentrations of molecular-grade MgCl2, dATP, dCTP, dGTP, and dTTP.
  • AccuStart II Taq DNA Polymerase
  • SYBR Green I dye
  • Inert AccuVue dye
  • Proprietary enzyme stabilizers and performance-enhancing additives.
  • Titrated reference dye (if applicable).

Storage conditions

PerfeCTa SYBR Green SuperMix is stable for 1 year when stored in a constant temperature freezer at -20°C, protected from light. For convenience, it may be stored unfrozen at 4°C for up to 6 months, protected from light. After thawing, mix thoroughly before using. Stabilized reagent demonstrates no functional loss of performance after 20 freeze-thaw cycles or 2 months at 20°C..

Resources

• 95068 PerfeCTa SYBR Green Supermix,UNG Protocol
• 95068 PerfeCTa SYBR Green Supermix,UNG MSDS
• SYBR SuperMixes and FastMixes Product Brochure

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