Luna® Universal qPCR Master Mix

Rapid, sensitive and precise dye-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)
    Cannot order from your location. Contact NEB for order information
  • 2 X
    500 rxn (5 x 1 ml)
    Cannot order from your location. Contact NEB for order information
  • 2 X
    1,000 rxn (10 x 1 ml)
    Cannot order from your location. Contact NEB for order information
  • 2 X
    2,500 rxn (1 x 25 ml)
    Cannot order from your location. Contact NEB for order information
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
     Dye-based quantitative PCR (qPCR) uses real-time fluorescence of a double-stranded DNA (dsDNA) binding dye, most commonly SYBR® Green I, to measure DNA amplification during 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.

    The NEB Luna Universal qPCR Master Mix is an optimized 2X reaction mix for real-time qPCR detection and quantitation of target DNA sequences using the SYBR®/FAM channel of most real-time qPCR instruments. 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 with fluorescent dyes 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 and DNA template. Genomic DNA or cDNA of interest can be quantitated with Luna qPCR, and existing as well as commercial qPCR assay primer sequences can be used.

    Figure 1: NEB’s Luna Universal qPCR Master Mix offers exceptional sensitivity, reproducibility and qPCR performance
    qPCR targeting human GAPDH was performed using the Luna Universal 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 qPCR Master Mix provides sensitive and accurate detection and quantitation across a wide variety of DNA sources
    The Luna Universal qPCR Master Mix is compatible with a broad range of genomic DNA sources. qPCR targets were quantitated with 50 ng – 0.5 pg genomic DNA as input using an ABI 7500 Fast real-time instrument. Genomic DNA was purified by typical column-based methods. In these examples, strong performance can be observed in the amplification of ACTB (encoding β-actin) from Mouse kidney genomic DNA, psbB (Photosystem II CP47 reaction center protein PsbB) from Tobacco, and RDN18 (18S ribosomal RNA) from Yeast.

    Figure 3: Extensive performance evaluation of commercially available dye-based qPCR reagents demonstrates the robustness and specificity of Luna
    qPCR reagents from NEB and other manufacturers were tested across 16–18 qPCR targets varying in abundance, length and %GC, using either Jurkat genomic DNA or Jurkat-derived cDNA as input (10 genomic DNA targets and 8 cDNA targets on Bio-Rad real-time instrument, 9 genomic and 7 cDNA targets on ABI instrument). For each testing condition, data was collected by 2 users and according to manufacturer’s 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: Bio-Rad, SsoAdvanced™ Universal SYBR® Green Supermix; Roche, FastStart™ SYBR Green Master; QIAGEN, QuantiTect® SYBR Green PCR Kit; ABI, PowerUP™ SYBR Green Master Mix; Promega®, GoTaq® qPCR Master Mix. NEB’s Luna Universal 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
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