Luna® Universal Probe qPCR Master Mix

Rapid, sensitive and precise probe-based qPCR detection and quantitation of target DNA and cDNA sequences.

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

Ordering Information

  • 2 X
    200 rxn (2 x 1 ml)
  • 2 X
    500 rxn (5 x 1 ml)
  • 2 X
    1,000 rxn (10 x 1 ml)
  • 2 X
    2,500 rxn (1 x 25 ml)
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

    Probe-based quantitative PCR (qPCR) uses real-time fluorescence released 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 significantly detectable 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. 

    The NEB Luna Universal Probe qPCR Master Mix is a 2X reaction mix optimized for real-time qPCR detection and quantitation of target DNA sequences using hydrolysis probes. It contains Hot Start Taq DNA Polymerase and has been 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). It also features dUTP for carryover prevention and a non-fluorescent, visible dye to monitor reaction setup. This dye does not spectrally overlap fluorophores commonly used for qPCR and will not interfere with real-time detection.

    The master mix formulation is supplied at 2X concentration and contains all PCR components required for amplification and quantitation of DNA except primers/probes and DNA template. Genomic DNA or cDNA of interest can be quantitated with Luna qPCR and existing as well as commercial qPCR assay primer/probe sequences can be used.

    Figure 1: NEB’s Luna Universal Probe qPCR Master Mix offers exceptional sensitivity, reproducibility and qPCR performance 
    qPCR targeting human GAPDH was performed using the Luna Universal Probe qPCR Master Mix over a 6-log range of input template concentrations (20 ng – 0.2 pg Jurkat-derived cDNA) with 8 replicates at each concentration. cDNA was generated from Jurkat total RNA using the NEB Protoscript® II First Strand cDNA Synthesis Kit (NEB #E6560).

    Figure 2: NEB’s Luna Universal Probe qPCR Master Mix offers robust performance in multiplex applications
    Singleplex (left) and multiplex (right) qPCRs targeting human GAPDH, ribosomal protein L32g and PI-3-Kinase-Related Kinase SMG1 were performed using the Luna Universal Probe qPCR Master Mix over a 5-log range of input template concentrations (20 ng – 2 pg Jurkat-derived cDNA) with 4 replicates at each concentration. 0.4 µM primer was used for the lower-copy target (SMG1) and 0.2 µM primer for each higher-copy target (L32g and GAPDH), in both multiplex qPCR (to account for copy number differences) and singleplex qPCR (to allow direct comparison).

    Figure 3: Extensive performance evaluation of commercially available probe-based qPCR reagents demonstrates the robustness and specificity of Luna
    Luna DNA probe
    qPCR reagents from NEB and other manufacturers were tested on 10 qPCR targets, varying in abundance, length and % GC, using either Jurkat genomic DNA or Jurkat-derived cDNA as input (5 targets each). For each testing condition, data was collected by 2 users and according to manufacturer specifications. 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 major manufacturers are shown: QIAGEN, QuantiTect® Probe PCR Kit; Bio-Rad, SsoAdvanced Universal Probes Supermix; Roche, FastStart® TaqMan® Probe Master; ABI, TaqMan Fast Advanced Master Mix; Promega®, GoTaq® Probe qPCR Master Mix. NEB’s Luna Universal Probe qPCR Master Mix outperformed all other reagents tested.  

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

    Product Categories:
    Luna® qPCR & RT-qPCR Products,
    PCR, qPCR & Amplification Technologies Products
    DNA Amplification, PCR and qPCR
    • Properties and Usage
    • Related Products
    • Notes
  • FAQs & Tech Tips
  • Protocols & Manuals
  • Other Tools & Resources
  • Quality & Safety
    • Quality Control Assays
    • Specifications
    • Certificate of Analysis
    • Safety Data Sheets
  • Legal Information
    • Legal And Disclaimers
Loading Spinner