Are there any performance changes in real-time PCR supermixes updated with the new ROX dye?

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Normalization of Real-Time PCR Fluorescence Data with ROX Passive Reference Dye


Question:

Are there any performance changes in real-time PCR supermixes updated with the new ROX dye?


Answer:

The Bio-Rad R&D team validated these updated supermixes with an extensive set of qPCR assays, evaluating relative performance, PCR efficiency, dynamic range, linearity, sensitivity, and specificity to ensure that the supermixes with the new dye provided the same high performance as the original formulations. Although the vast majority of data, in terms of absolute Cq values, were identical, we noted a few assays that demonstrated Cq shifts ±0.5. This shift does not impact qPCR assay results in any manner, as Cq values in and of themselves are meaningless values.

See Normalization of Real-Time PCR Fluorescence Data with ROX Passive Reference Dye in our Applications and Technologies pages to learn why passive reference dyes are required for data normalization in some real-time PCR systems and not others. Although Bio-Rad real-time PCR detection systems do not require ROX for internal normalization, Bio-Rad’s universal real-time PCR supermixes are designed for high performance on all real-time PCR detection systems, both systems requiring ROX normalization and ROX-independent systems.

View our Understanding Real-Time Supermixes tutorial to learn more about qPCR data analysis and how to compare supermix performance.


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