Luna® Universal Probe One-Step RT-qPCR Kit

Rapid, sensitive and precise probe-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

Ordering Information

$name
  • 200 reactions
    $211.00
  • 500 reactions
    $475.00
  • 1,000 reactions
    $833.00
  • 2,500 reactions
    $1,833.00
Did you know this product can be customized or purchased in larger volumes? Submit an inquiry to find out more about customization options.
  • Product Information
    Sample
     
     The NEB Luna Universal Probe One-Step RT-qPCR Kit is optimized for real-time quantitation of target RNA sequences using hydrolysis probes. 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. Probe-based qPCR/RT-qPCR monitors an increase in fluorescence upon 5´ → 3´ exonuclease cleavage of a quenched, target-specific probe to measure DNA amplification at each cycle of a PCR. 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.

    In the Luna Universal One-Step Probe 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 Probe One-Step Reaction Mix is supplied at 2X concentration and contains Hot-Start Taq DNA Polymerase, dNTPs, 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 fluorophores commonly used in qPCR 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.

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    Figure 1: NEB’s Luna Universal Probe One-Step RT-qPCR Kit offers exceptional sensitivity, reproducibility and RT-qPCR performance
    RT-qPCR targeting human GAPDH was performed using the Luna Universal Probe One-Step RT-qPCR Kit over an 8-log range of input template concentrations (1 μg – 0.1 pg Jurkat total RNA) with 8 replicates at each concentration. Reaction setup and cycling conditions followed recommended protocols, including a 10-minute RT step at 55°C for the thermostable Luna WarmStart Reverse Transcriptase.


    Figure 2: NEB’s Luna Universal Probe One-Step RT-qPCR Kit offers robust performance in multiplex applications
    Luna
    Multiplex RT-qPCR targeting human GAPDH, ribosomal protein L32g and PI3-Kinase-Related Kinase SMG1 was performed using the Luna Universal Probe One-Step RT-qPCR Kit over a 7-log range of input template concentrations (1 μg – 1 pg Jurkat total RNA) with 4 replicates at each concentration. Amplification plots are shown both overlayed (left) and for each multiplex target (right). To account for copy number differences, 0.4 µM primer was used for lower-copy target (SMG1) and 0.2 µM primer for higher-copy targets (L32g and GAPDH). Luna maintains superior efficiency, reproducibility, sensitivity and performance in multiplex RT-qPCR.



    Figure 3: Extensive performance evaluation of commercially available probe-based RT-qPCR reagents demonstrates the robustness and specificity of Luna
    Luna
    Commercially-available RT-qPCR reagents were tested on 7 RT-qPCR targets varying in abundance, length, and %GC. Data was collected by 2 users and according to manufacturer’s recommendations. Results were evaluated for efficiency, low input detection and lack of non-template amplification (where ΔCq = average Cq of non-template control – average Cq of lowest input). In addition, consistency, reproducibility and overall curve quality were assessed (Quality Score). Bar graph indicates % of targets that met acceptable performance criteria (indicated by green box on dot plot and Quality Score > 3). Results for NEB and other suppliers are shown: Quanta, qScript XLT 1-Step RT-qPCR ToughMix®; ABI, TaqMan® RNA-to-Ct 1-Step Kit; QIAGEN, QuantiFast® Probe RT-PCR Kit; Bio-Rad, iTaq Universal Probes One-Step Kit; Promega®, GoTaq® Probe 1-Step RT-qPCR System. NEB’s Luna Universal Probe One-Step RT-qPCR Kit outperformed all other reagents tested.

    Learn more about our comprehensive qPCR/RT-qPCR testing and “dots in boxes” data visualization.

    Kit Components

    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 Probe One-Step Reaction Mix -20 2 X
    Product Categories:
    Luna® qPCR & RT-qPCR Products,
    PCR, qPCR & Amplification Technologies Products
    Applications:
    PCR,
    DNA Amplification, PCR and qPCR
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