Luna® Universal One-Step RT-qPCR Kit
Rapid, sensitive and precise dye-based qPCR detection and quantitation of target RNA targets
Make a simpler choice
- One product per application simplifies selection
- Convenient master mix formats and user-friendly protocols simplify reaction setup
- Non-interfering, visible tracking dye helps to eliminate pipetting errors
Experience best-in-class performance
- All Luna® products have undergone rigorous testing to optimize specificity, sensitivity, accuracy and reproducibility
- Products perform consistently across a wide variety of sample sources
- A comprehensive evaluation of commercially-available qPCR and RT-qPCR reagents demonstrates superior performance of Luna products
Optimize your RT-qPCR with Luna WarmStart® Reverse Transcriptase
- Novel, thermostable reverse transcriptase (RT) improves performance
- WarmStart RT paired with Hot Start Taq increases reaction specificity and robustness
“Dots in Boxes" Visualization of qPCR Data
Overview of qPCR
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Rapid, sensitive and precise dye-based qPCR detection and quantitation of RNA targets.The NEB Luna Universal One-Step RT-qPCR Kit is optimized for dye-based real-time quantitation of target RNA sequences via the SYBR®/FAM fluorescence channel of most real-time instruments. Dye-based qPCR/RT-qPCR uses real-time fluorescence of a double-stranded DNA (dsDNA) binding dye, most commonly SYBR Green I, to measure DNA amplification after each PCR cycle. At a point where the fluorescence signal is confidently detected over the background fluorescence, a quantification cycle or Cq value can be determined. Cq values can be used to evaluate relative target abundance between two or more samples, or to calculate absolute target quantities in reference to an appropriate standard curve derived from a series of known dilutions.
One-Step RT-qPCR provides a convenient and powerful method for RNA detection and quantitation. In a single tube, RNA is first converted to cDNA by a reverse transcriptase, and then a DNA-dependent DNA polymerase amplifies the cDNA, enabling quantitation via qPCR.
In the Luna One-Step RT-qPCR Kit, Hot Start Taq DNA Polymerase is combined with a novel WarmStart-activated reverse transcriptase, allowing dual control of enzyme activity via reversible, aptamer-based inhibition. This temperature-dependent activation helps to prevent undesirable non-specific priming and extension prior to thermocycling, providing added security for setting up reactions at room temperature. The engineered WarmStart Luna Reverse Transcriptase also possesses higher thermostability than many other RTs, allowing an optimal reaction temperature of 55°C. For difficult targets/templates, higher RT step temperatures of up to 60°C can be used without compromising Luna performance.
Note that to ensure full activation of the WarmStart Luna RT, incubation at temperatures lower than 50°C is not recommended.
The Luna Universal One-Step Reaction Mix is supplied at 2X concentration and contains Hot-Start Taq DNA Polymerase, dNTPs, a fluorescent dsDNA-binding dye, and all required buffer components. It is formulated with a unique passive reference dye that is compatible across a variety of instrument platforms, including those that require a high or low ROX reference signal. The Reaction Mix also features dUTP for carryover prevention and a non-fluorescent visible dye for monitoring reaction setup. This visible dye does not overlap spectrally with the included dsDNA-binding dye and does not interfere with real-time detection.
The Luna WarmStart RT Enzyme Mix is supplied at 20X concentration and contains Luna WarmStart Reverse Transcriptase as well as Murine RNase Inhibitor to aid in preventing RNA degradation (see also template preparation in product manual ). It is compatible with various RNA sample types (total RNA, poly(A)-RNA, etc.) and sources.
The following reagents are supplied with this product:
Store at (°C) Concentration Nuclease-free Water -20 Luna® WarmStart® RT Enzyme Mix -20 20 X Luna® Universal One-Step Reaction Mix -20 2 X
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