PerfeCTa qPCR ToughMix

PerfeCTa qPCR ToughMixes enable expression analysis and other quantitative applications from crude samples such as lysates, blood spots, plant tissues and clinical samples.

Features & Benefits:

  • Bullet-Proof PCR – easily overcome tough PCR inhibitors
  • Short Run Times – fast cycling protocols enable more experiments
  • Optical Clarity Technology – high precision in reduced reaction volumes
  • Repeatable results – hot-start polymerase highly stable at ambient temperature
  • ToughMix additives neutralize PCR inhibitors to ensure reliable assay performance with a spectrum of starting materials including: clinical specimens, plants, soil, environmental or complex food matrices.
  • Superior assay sensitivity and target precision with AccuStart™ II enzyme technology – maximum-yielding, engineered Taq DNA polymerase with stringent, ultrapure antibody hotstart.
  • Easy-to-use 2X concentrated master mixes with AccuVue™ plate loading dye and optimized passive reference dye for simplified reaction setup.
  • Supports efficient vortex mixing with proprietary anti-foaming technology.
  • Consistent and reliable performance with exceptional reagent stability (uninhibited after 20X freeze-thaw or 30 days at 22°C).

PerfeCTa qPCR ToughMix is a 2X concentrated ready-to-use reaction cocktail for PCR amplification of DNA templates that relieves several types of PCR inhibition commonly encountered with crude extracts, environmental specimens, plant tissues, animal tissues, and complex food matrices. This robust real-time qPCR reagent provides maximum sensitivity and PCR efficiency with a variety of fluorogenic probe chemistries, including TaqMan® hydrolysis probes. The only user-supplied components are primers, probe(s), and DNA template. Pre-blended with inert AccuVue plate loading dye to help minimize pipette errors during setup and provides visual confirmation of thorough mixing. A key component of PerfeCTa qPCR ToughMix is an ultra pure, highly processive thermostable DNA polymerase that is combined with high avidity monoclonal antibodies. This proprietary polymerase mix is highly resistant to PCR inhibitors and provides an extremely stringent automatic hot-start allowing reaction assembly, and temporary storage, at room temperature prior to PCR amplification. PerfeCTa qPCR ToughMix delivers exceptional performance with either fast or conventional PCR cycling protocols. UNG containing versions are blended with Uracil N-glycosylase to eliminate potential post-PCR carryover contamination associated with routine molecular testing.

Comparison to Conventional Master Mixes

PerfeCTa qPCR ToughMix stands up to the challenge where other qPCR master mixes fall apart.  qPCR is performed on clean template and in the presence of PCR inhibitors with PerfeCTa ToughMix and three relevant competitor’s Master Mixes (Figure 1). Average CT values obtained with PerfeCTa ToughMix are unaffected by the presence of the PCR inhibitors while competitor’s mixes show signs of inhibition and in some cases fail to amplify.

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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
  • BioRad iCycler iQ™
  • BioRad MyiQ™
  • BioRad iQ™5

Contents

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

Storage conditions

PerfeCTa qPCR ToughMix is stable for 2 years when stored in a constant temperature freezer at or below -20°C. For convenience, it may be stored unfrozen at 4°C for up to 6 months. After thawing, mix thoroughly before using. Repeated freezing and thawing does not affect PCR performance.

Resources

Ordering

Cat# 95112-250 250 x 20 µl rxns (2 x 1.25 mL)
Cat# 95112-012 1250 x 20 µl rxns (10 x 1.25 mL)
Cat# 95112-05K 5000 x 20 µl rxns (1 x 50 mL)