Description
GoldBio’s Goof-Proof™ qPCR Master Mix is a fast, sensitive alternative to SYBR®-based mixes, using EvaGreen® dye for enhanced signal clarity and low background fluorescence.
This 2X hot-start master mix is designed for real-time PCR applications where performance and error-free setup are essential. Goof-Proof™ is a mistake-prevention master mix which contains a low concentration of blue dye. Additionally, Goof-Proof™ includes a DNA template buffer which has a higher concentration of blue dye. With the addition of the 2X Goof-Proof™ Master Mix to the reaction, it will appear light blue. And when you add the template, containing DNA template buffer, the color will turn darker blue.
Goof-Proof™ allows you to see quickly and easily whether you forgot to add the master mix or the template to any of your reactions, preventing pipetting mistakes and wasted time, reagents and money. The 2X Goof-Proof™ Master Mix can also be used without the Template Buffer, if desired.
Its EvaGreen® offers superior safety, higher sensitivity, and sharp melt curves, making Goof-Proof™ ideal for gene expression analysis, low-abundance target detection, and high-throughput qPCR workflows. With a built-in pipetting dye and ROX-free formulation, Goof-Proof™ delivers consistent, publication-ready results for any lab seeking safer, smarter qPCR performance.
Note: We offer Goof-Proof™ qPCR Master Mix with Rox (Catalog # G-705) which includes a separate vial of ROX Reference dye, and instructions for how much ROX to add depending on which protocol and instrument you are using.
Kit components
- Goof-Proof™ 2X Master Mix -
The 2X Master Mix contains GoldBio HotStart Taq DNA Polymerase, dNTPs and a low concentration of an inert blue dye. The dye helps the user distinguish between empty wells and wells containing master mix.
- Goof-Proof™ Template Buffer -
The 40X Template Buffer contains a high concentration of inert blue dye. Adding the template buffer to the reaction mix makes it "Goof-Proof" to track which wells have had template added to the PCR reactions. Template Buffer is not required for the Master Mix to function, but is designed to help alleviate mistakes with template addition.
Kit Information
Dye System: EvaGreen® dye (non-toxic, high signal-to-noise, probe-free detection)
Enzyme: Hot-start DNA polymerase for high specificity and minimal background
Visual Tracking: Blue dye included for pipetting confirmation
Application Compatibility: Works with most real-time PCR instruments that do not requiring ROX
Common Applications:
(Click each for more information)
High-Precision Real-Time PCR with EvaGreen® Dye
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Purpose: To amplify and quantify DNA targets using real-time detection with a safer, brighter alternative to SYBR® Green.
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How It Works: EvaGreen® dye binds to double-stranded DNA and fluoresces without inhibiting polymerase activity, offering high signal-to-noise ratios with reduced nonspecific amplification.
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Applications: Gene expression analysis, quantification of genomic targets, validation of NGS data.
Mao, F., et al. (2007). Characterization of EvaGreen and the implication of its physicochemical properties for qPCR applications. BMC Biotechnology, 7, 76.
Eco-Friendly qPCR for Routine Lab Use
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Purpose: To perform environmentally safer and less toxic qPCR experiments without compromising sensitivity or precision.
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How It Works: EvaGreen® is non-mutagenic, non-cytotoxic, and safer for waste disposal than ethidium bromide or SYBR® Green.
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Applications: Green labs, teaching environments, high-throughput qPCR facilities.
Mao, F., et al. (2007). Characterization of EvaGreen and the implication of its physicochemical properties for qPCR applications. BMC Biotechnology, 7, 76.
Quantitative PCR for Low-Abundance or Rare Transcripts
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Purpose: To sensitively detect low-copy or poorly expressed genes in biological samples.
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How It Works: Goof-Proof™’s formulation includes a hot-start polymerase and EvaGreen®, enabling high-specificity amplification with bright fluorescence and low background.
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Applications: Biomarker discovery, pathogen detection, expression profiling.
Kubista, M., et al. (2006). The real-time polymerase chain reaction. Molecular Aspects of Medicine, 27(2–3), 95–125.
Melt Curve Analysis for Specificity Assessment
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Purpose: To verify qPCR specificity and detect primer-dimer formation without the use of probes.
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How It Works: EvaGreen® dye produces sharp, reliable melt curves that distinguish target amplicons from nonspecific products or primer-dimers.
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Applications: Primer validation, qPCR troubleshooting, multiplex optimization.
Mao, F., et al. (2007). Characterization of EvaGreen and the implication of its physicochemical properties for qPCR applications. BMC Biotechnology, 7, 76.
Varga, A., & James, D. (2006). Real-time RT-PCR and SYBR Green I melting curve analysis for the identification of Plum pox virus strains C, EA, and W. Journal of Virological Methods, 132(1–2), 146–153.
Fast, Visual Reaction Assembly for Error Prevention
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Purpose: To reduce setup mistakes and streamline pipetting, especially in 96- and 384-well formats.
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How It Works: Goof-Proof™ includes a visible tracking dye that confirms reagent addition, reducing the risk of missed wells and improving reproducibility.
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Applications: High-throughput qPCR, teaching labs, assay development.
Taylor, S. C., et al. (2019). The ultimate qPCR experiment: producing publication-quality, reproducible data the first time. Trends in Biotechnology, 37(7), 761–774.
Key Benefits:
Visual setup verification
Built-in pipetting dye reduces missed wells and improves reproducibility.
Probe-free detection with melt curve validation
Ideal for applications where specificity and simplicity matter
ROX-free flexibility
Compatible with a wide range of real-time PCR instruments that do not require normalization.
Storage/Handling
Store the kit at -20°C.
Product may be shipped on blue ice without reducing performance. Please place at the recommended storage conditions upon receipt of the product.
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