This document describes how to avoid generating nonspecific amplification when using TaqMan 'Assays-on Demand' gene expression products.
This document addresses the issue of low amplification efficiency and nonspecific PCR products, such as primer-dimers, which can lead to erroneous data analysis in real-time PCR. It highlights that pre-designed assays, like Assays-on-Demand (AOD), can sometimes generate these artifacts. The document presents data comparing Agilent's Brilliant III Ultra-Fast reagents against a competitor's fast QPCR master mix.
| Amplification Efficiency (GUS) | 94.8% |
|---|---|
| Amplification Efficiency (TBP) | 99.7% |
| Correlation Coefficient (GUS) | 0.999 |
| Correlation Coefficient (TBP) | 0.998 |
The novel hot start technology of Brilliant III prevents the generation of non-specific secondary products like primer-dimers.
Primer-dimers compete with specific product amplification, reducing efficiency, sensitivity, and dynamic range.