Luna® Direct Probe qPCR 4X Master Mix with UDG (No ROX)
The Luna Direct Probe qPCR 4X Master Mix with UDG (No ROX) enables robust qPCR performance from challenging sample types and crude lysates. Enhanced inhibitor tolerance and a concentrated 4X formulation support higher sample input for improved assay sensitivity..
For instruments that do not require the ROX passive reference dye
Enhanced inhibitor tolerance delivers reliable amplification from both crude and purified samples
Accelerate qPCR workflows with extension times as short as 10 seconds
Increase assay sensitivity with a 4X formulation that supports higher sample input
Maximize your throughput by multiplexing up to 6 targets
Hot Start Taq increases reaction specificity and robustness, enabling room temperature setup
Reduce the risk of carryover contamination with thermolabile UDG and dUTP included in the mix
Eliminate pipetting errors with non-interfering, visible blue tracking dye
The Luna Direct Probe qPCR 4X Master Mix with UDG (No ROX) enables robust qPCR performance from challenging sample types and crude lysates, combining enhanced inhibitor tolerance with a concentrated 4X formulation that supports higher sample input for improved assay sensitivity. It supports multiplex detection of up to 3 gDNA targets or 6 cDNA targets in a single reaction. The mix contains Hot Start Taq DNA Polymerase, dNTPs, and Murine RNase Inhibitor in an optimized buffer. Hot Start Taq minimizes non-specific amplification, improves reaction specificity, and allows for room temperature setup.
Thermolabile UDG and dUTP help prevent carryover contamination. The thermolabile UDG is fully inactivated above 50°C, eliminating any impact on qPCR performance.
This formulation contains no ROX reference dye and is compatible with any instrument that does not require ROX.
Luna qPCR (No ROX) product comparison
Figure 1: Multiplex detection (6 targets) with the Luna Direct Probe qPCR 4X Mix with UDG
Multiplex qPCR was performed using the Luna Direct Probe qPCR 4X Mix with UDG over a 5-log range of LunaScript® RT SuperMix–synthesized cDNA (20 ng to 2 pg) on a qTOWERiris 96 touch UV ready real-time instrument. Amplification standard curves and efficiencies for each of the six human targets are shown. Reactions (20 µl) contained primers and probes at 200 nM each for GAPDH, SDHA, and APD; 100 nM each for ACTIN and RPL32; and 400 nM each for SMG1. The reactions were run under the product-recommended cycling conditions. All six targets were detected linearly in multiplex format, demonstrating strong amplification efficiency and high R² values.
Figure 2: Reliable amplification from multiple purified and crude sample types
Reliable amplification from multiple purified and crude sample types. qPCR targeting Heme from human blood, tet(o) from wastewater, 18S from corn and bacterial 16S from human feces was performed using the Luna Direct Probe qPCR 4X Master Mix with UDG. Performance with purified and crude samples in 20 μl reactions was evaluated.
Purified samples: 50 ng–50 pg DNA from K2-EDTA Human blood; 300 ng–30 pg DNA from wastewater; 50 ng–5 pg DNA from corn; 100 ng–10 pg DNA from human feces.
Direct samples: 10, 15 and 20% K2-EDTA Human blood; 10, 20 and 40% composite wastewater; 2.5% corn leaf lysate; 5% human feces lysate.
Sensitive, robust performance can be observed across a 5-log dilution using purified nucleic acid from various sample types. In addition, Luna Direct robustly amplifies targets directly from crude sample types. NTC amplification was either not observed or at least 8 Cq values higher than the Cq of the lowest sample input.
Figure 3: Luna Direct enables robust cross-platform performance of a multiplex STI assay
Multiplex STI qPCR assay targeting Chlamydia trachomatis (ompA), Neisseria gonorrhoeae (porA), Trichomonas vaginalis (TV-E650 repeat), and human RNase P (RPPH1) was performed using Luna Direct Probe qPCR 4X Master Mix with UDG in 20 μl reactions across various real-time PCR instruments. Assay performance was evaluated using a 5-log dilution series (10–100,000 copies per reaction) of quantitative genomic DNA from C. trachomatis (ATCC® VR-885DQ™), N. gonorrhoeae (ATCC VR-19424DQ™), and T. vaginalis (ATCC VR-30001DQ) in the presence of 5 ng Jurkat genomic DNA (BioChain®, #D1255815). Consistent, sensitive amplification was observed for each target across all platforms, demonstrating robust multiplex performance over a broad dynamic range.
A non-fluorescent visible blue tracking dye is included to assist in pipetting into clear vessels. The tracking dye does not overlap spectrally with fluorophores commonly used in qPCR and does not interfere with real-time detection.
The Luna Direct Probe qPCR 4X Master Mix with UDG (No ROX) contains an inert visible blue tracking dye to eliminate pipetting errors
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