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All NEB products undergo rigorous testing to ensure optimal performance, and Luna is no exception. We took into consideration numerous important traits when evaluating qPCR, including specificity, sensitivity, accuracy and reproducibility, to develop best-in-class qPCR products. Furthermore, we did a comprehensive evaluation of many commercially-available qPCR and RT-qPCR products, and developed a method of analysis that allows you to quickly compare and evaluate the performance of these products. We wanted to be sure that Luna products will perform to your expectations for all of your targets.

Benefit from rigorous testing

  • All Luna reagents were evaluated for specificity, sensitivity, efficiency, accuracy, linearity and reproducibility, and optimized for performance
  • Development of novel “dots in boxes” analysis method for concise visual presentation of comprehensive evaluations

Use with ALL your targets

  • Extensive panel testing and evaluation ensures reproducible results
  • Luna reagents are optimized for strong performance across a wide range of targets and templates, including those of varied length, GC-content and abundance

Videos

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LUNA qPCRLearn the basics of qPCR in this short animation.

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Evaluating qPCR results: capturing performance as “dots in boxes”

NEB has developed a method to better evaluate the large amount of qPCR data generated in an experiment. The output of this analysis is known as a dot plot, and captures the key features of a successful, high-quality qPCR experiment as a single point. This method of analysis allows many targets and conditions to be compared in a single graph.

For each experiment, triplicate reactions are set up across a five-log range of input template concentrations (Amplification plot, bottom-left). Three non-template control (NTC) reactions are also included, for a total of 18 reactions per condition/target. Efficiency (%) is calculated (Standard curve, top-left) and is plotted against ΔCq (dot plot, center), which is the difference between the average Cq of the lowest input and the NTC. This parameter captures both detection of the lowest input and non-template amplification. Acceptable performance criteria are defined as an Efficiency of 90-110% and a ΔCq of ≥ 3 (green box).

Other performance criteria are captured using a 5-point Quality Score (top-right). Included are:

  1. 1. Linearity of amplification, as indicated by the R2 standard curve
  2. 2. Reproducibility, as indicated by the consistency of triplicate Cq values for each input concentration
  3. 3. Fluorescence consistency, as indicated by similar endpoint fluorescence (RFUmax)
  4. 4. Curve steepness
  5. 5. Sigmoid curve shape

Quality Score is represented by the size and fill of the plotted dot, with experiments that pass all performance criteria represented by a solid dot within the box. These scoring methods were built upon the MIQE qPCR/RT-qPCR guidelines (Bustin, S.A. et al. (2009) Clin Chem. 55, 611-22 and Trombley Hall, A. et al. (2013) PLoS One 8(9):e73845). An example of this analysis is shown below.

Breaking it down: how we translate qPCR data into “dots in boxes”

See how Luna products stack up

Comprehensive market-wide evaluation of qPCR and RT-qPCR products reveals that Luna outperforms the competition.

Evaluation of commercially-available qPCR products demonstrates robustness and specificity of Luna

Available products

PRODUCTS REF. SIZE PRICE MORE INFO
NEW LunaScript® RT Master Mix Kit (Primer-free) E3025S/L 25/100 rxns In den Shop wechseln Information Produit
NEW Luna SARS-CoV-2 RT-qPCR Multiplex Assay E3019S/L 96/480 rxns In den Shop wechseln Information Produit
NEW Luna Probe One-Step RT-qPCR 4x Mix with UDG M3019S/L/X/E 200/500/1000/2000 rxns In den Shop wechseln Information Produit
Luna Cell Ready One-Step RT-qPCR Kit E3030S 100 rxns In den Shop wechseln Information Produit
Luna Cell Ready Probe One-Step RT-qPCR Kit E3031S 100 rxns Boutique en ligne Information Produit
Luna Cell Ready Lysis Module E3032S 100 rxns Boutique en ligne Information Produit
Luna Universal qPCR Master Mix M3003S 200 rxns (2x1ml) In den Shop wechseln Produktinformationen
M3003L 500 rxns (5x1ml) In den Shop wechseln Produktinformationen
M3003X 1000 rxns (10x1ml) In den Shop wechseln Produktinformationen
M3003E 2500 rxns (1x25ml) In den Shop wechseln Produktinformationen
Luna Universal Probe qPCR Master Mix M3004S 200 rxns (2x1ml) In den Shop wechseln Produktinformationen
M3004L 500 rxns (5x1ml) In den Shop wechseln Produktinformationen
M3004X 1000 rxns (10x1ml) In den Shop wechseln Produktinformationen
M3004E 2500 rxns (1x25ml) In den Shop wechseln Produktinformationen
Luna Universal One-Step RT-qPCR Kit E3005S/L/X/E 200/500/1000/2500 rxns In den Shop wechseln Produktinformationen
Luna Universal Probe One-Step RT-qPCR Kit E3006S/L/X/E 200/500/1000/2500 rxns In den Shop wechseln Produktinformationen
LunaScript RT SuperMix Kit E3010S/L 25/100 rxns In den Shop wechseln Produktinformationen

Further information can be found in our Technical Resources section or at neb.com