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This learning center is your guide to the theory and implementation of the qPCR technique as it is used in gene expression analysis, beginning with RNA iQ™ SYBR® Green Supermix Manual Guidelines for Optimizing qPCR Reactions . Figure 1. SYBR® Green I DNA-binding in a real-time PCR reaction.Always use dedicated pipets for qPCR. 4. Use aerosol-resistant filter tips. 5. Use PCR-grade water. 6. Clean bench with 10% bleach, not ethanol! Real-time PCR can be used for both qualitative and quantitative analysis; choosing the best method for your Real-time qPCR Workflow Selection Guide. Learn to design, optimize, and validate real-time PCR assays. Primer and probe design, target selection, gradient, melt curve, and multiplexing for qPCR It includes guidelines for designing the best real-time PCR assay for your experiments and explains how real-time PCR data are used in various applications. Reagent Comparison Guide for Real-Time PCR. Visit us on the Web at bio-rad.com. 4. Reagent and Assay Comparison Matrix. Preparation of. Samples and. Perform one-step multiplex RT-qPCR with minimal cross-reactivity using Reliance One-Step Multiplex Supermix and. PrimePCR Probe Assays expertly designed for This guidebook comes with an easy-to-review real-time PCR quick guide to use at the bench. The quick guide covers the fundamentals of qPCR, design, optimization