Testing shows that TaqMan Assays and master mix outperform the equivalent Roche reagents on the Roche LightCycler 480 II qPCR system
We conducted side-by-side tests running Applied Biosystems TaqMan Gene Expression Assays with TaqMan Fast Advanced Master Mix and TaqPath 1-Step Multiplex Master Mix vs Roche LightCycler® Probes Master and Multiplex Virus Master on the Roche LightCycler 480 qPCR system. In both series of tests we looked for reproducibility and dynamic range, as well as singleplex vs duplex/4-plex performance. The results are in—and they just might surprise you.
Do you currently use the Roche Universal Probe Library (UPL)?
You probably already know that Roche is discontinuing their UPL product line. Did you also know that TaqMan Assays and master mix work great on Roche instruments and might just provide even better performance than what you’ve been experiencing? Check out the results from these tests, then check out TaqMan solutions for your research.
TaqMan Gene Expression Assays with TaqMan Fast Advanced Master Mix showed greater PCR efficiency when compared to Roche reagents
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Figure 1. TaqMan Fast Advanced Master Mix demonstrated broad linear dynamic range and reliable quantification at low copy numbers. Amplification plots from an 8-log dilution series of 50 ng/µL human cDNA amplified in five replicate reactions with an 18S rRNA TaqMan Gene Expression Assay on a Roche LightCycler 480 II Real-Time PCR Detection System. LightCycler 480 Probes Master (left) was compared to TaqMan Fast Advanced Master Mix (right). Note the more consistent results with TaqMan Fast Advanced Master Mix at the lowest concentration.
TaqMan Gene Expression Assays with TaqPath 1-Step Multiplex Master Mix delivered higher sensitivity and efficiency than the Roche LightCycler reagents—with no loss of performance when multiplexing
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Figure 2. Comparison of singleplex vs 4-plex performance of Roche LightCycler Multiplex RNA Virus and TaqPath 1-step Multiplex Master Mix (no ROX) on a Roche LightCycler 480 II Real-Time PCR Detection System. Thirty-two TaqMan Gene Expression Assays were run in both 4-plex (5 replicates) and in simplex (3 replicates). The differences in Cq (Δ Average Cq) and slope of the standard curve (Δ Slope) between the assays run in 4-plex as compared to simplex are plotted here. Note the small Δ Average Cq and Δ Slope values with TaqPath 1-Step Multiplex Master Mix, indicating that there is no loss of performance when the reaction is run in 4-plex as compared to simplex.
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