Description
GoldBio’s Goof-Proof™ qPCR Master Mix with ROX is a high-performance master mix designed for fast, reliable, and error-resistant real-time PCR.
Formulated with hot-start DNA polymerase, optimized buffer, ROX passive reference dye, and a built-in visual pipetting indicator, Goof-Proof™ ensures accurate amplification, minimized background, and consistent Ct values, even in high-throughput formats.
This master mix is ideal for gene expression analysis, CRISPR validation, and multiplex workflows. It is instrument-compatible and publication-proven. Whether you're a beginner or an expert, Goof-Proof™ takes the guesswork out of qPCR every time.
The benefit of Goof-Proof™ is that it is a mistake-prevention master mix that 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.
Note: We also offer Goof-Proof™ qPCR Master Mix (Catalog # G-700) without ROX for instruments that do not require the ROX reference dye.
Kit components
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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.
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Goof-Proof™ Template Buffer -
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.
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ROX Reference Dye -
Proof™ qPCR Master Mix with ROX includes a vial of ROX reference dye. ROX is necessary for accurate Ct determination from well to well with certain instruments (such as some ABI and Stratagene PCR instruments, refer to your instrument instructions to see if ROX is required). ROX may add noise to melt curve analysis, which could be mistaken for real peaks. In the case of unexpected peaks, un-check “ROX” in the “Passive Reference Dye” box in the system software to prevent data collection from the ROX fluorescence channel and re-analyze the data.
Kit Information
Dye System: Includes ROX passive reference dye
Enzyme: Hot-start DNA polymerase for high specificity
Visual Tracking: Blue dye included for pipetting confirmation
Application Compatibility: Validated on most real-time PCR platforms
Common Applications:
(Click each for more information)
Quantitative PCR for Gene Expression Analysis
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Purpose: To measure relative or absolute gene expression levels in various biological samples using real-time PCR.
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How It Works: Goof-Proof™ qPCR Master Mix contains a hot-start DNA polymerase, dNTPs, ROX passive reference dye, and an optimized reaction buffer. This formulation ensures accurate fluorescence quantification on a cycle-by-cycle basis as amplification proceeds.
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Applications: Analysis of mRNA, non-coding RNA, or transgene expression across tissues or experimental conditions.
Bustin, S. A., et al. (2009). The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clinical Chemistry, 55(4), 611–622.
High-Specificity Detection of Low-Abundance Targets
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Purpose: To amplify rare transcripts or low-copy genes with minimal nonspecific products.
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How It Works: The hot-start polymerase and proprietary buffer chemistry reduce primer-dimer formation and nonspecific amplification, resulting in clean amplification curves and sharp melt profiles.
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Applications: Detection of rare mRNA isoforms, SNP genotyping, or microbial diagnostics.
Kubista, M., et al. (2006). The real-time polymerase chain reaction. Molecular Aspects of Medicine, 27(2–3), 95–125.
qPCR for CRISPR Validation and Editing Efficiency
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Purpose: To confirm gene editing events and quantify insertion or deletion frequencies following CRISPR-based genome editing.
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How It Works: Goof-Proof™ enables precise SYBR-based quantification using primers flanking the edited locus, allowing accurate measurement of on-target editing efficiency.
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Applications: Genomic deletion confirmation, knock-in screening, and off-target effect analysis.
Brinkman, E. K., et al. (2014). Easy quantitative assessment of genome editing by sequence trace decomposition. Nucleic Acids Research, 42(22), e168.
Fast and Reproducible qPCR in High-Throughput Formats
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Purpose: To accelerate reaction setup and reduce pipetting errors in 96- or 384-well qPCR workflows.
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How It Works: The 2X ready-to-use formulation includes visual tracking dyes to confirm proper reaction assembly while maintaining full qPCR compatibility.
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Applications: Plate-based screens, biomarker discovery, and time-course gene expression studies.
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:
Minimizes pipetting errors with visible tracking
Blue dye provides real-time visual confirmation of mix addition in each well.
High specificity and reproducibility
Reduces non-specific amplification and delivers consistent Ct values.
ROX normalization ensures instrument compatibility
Passive reference dye supports well-to-well signal normalization on ROX-enabled machines.
Streamlined for qPCR success
Pre-mixed, validated formulation accelerates setup and reduces variability.
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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