Item# Description List Price Web Price Qty
    E3005E Luna Universal One-Step RT-qPCR Kit - 2,500 rxns $2,887.00
    $2,598.30
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    E3005L Luna Universal One-Step RT-qPCR Kit - 500 rxns $748.00
    $673.20
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    E3005S Luna Universal One-Step RT-qPCR Kit - 200 rxns $332.00
    $298.80
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    E3005X Luna Universal One-Step RT-qPCR Kit - 1,000 rxns $1,299.00
    $1,169.10
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    *On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
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    Product Introduction

    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

    Product Information

    Description

    Sample
     

    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.


    Figure 1: NEB’s Luna Universal One-Step RT-qPCR Kit offers exceptional sensitivity, reproducibility and RT-qPCR performance
    RT-qPCR targeting human GAPDH was performed using the Luna Universal 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 One-Step RT-qPCR Kit provides sensitive and accurate detection and quantitation across a wide variety of RNA sources
    Luna
    The Luna Universal One-Step RT-qPCR Kit yields high-quality results across a broad range of RNA source organisms (mammals, plants, yeast and bacteria) and purification methods (commercial, column-purified or RNAlater-preserved total RNA; purified mRNA). RT-qPCR targets were quantitated with 50 ng – 5 pg total RNA as input using either ABI 7500 Fast or ABI QuantStudio® 6 real time instruments.


    Figure 3: Extensive performance evaluation of commercially available dye-based RT-qPCR reagents demonstrates the robustness and specificity of Luna
    Luna
    Commercially-available RT-qPCR reagents were tested across 24 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: QIAGEN, QuantiFast® SYBR® Green RT-PCR Kit; ABI, Power SYBR Green RNA-to-Ct 1-Step Kit; Bio-Rad, iTaq® Universal SYBR Green One-Step Kit; Promega®, GoTaq® 1-Step RT-qPCR System; Quanta, qScript One-Step SYBR Green RT-qPCR Kit. NEB’s Luna Universal 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 0 %
    Luna® WarmStart® RT Enzyme Mix -20 20 X
    Luna® Universal One-Step Reaction Mix -20 2 X
    Product Categories:
    Luna® qPCR & RT-qPCR Products,
    PCR, qPCR & Amplification Technologies
    Applications:
    PCR,
    RT-qPCR, RT-PCR and cDNA Synthesis,
    DNA Amplification, PCR & qPCR

    Properties and Usage

    Storage Temperature

    -20°C

    Faqs & Tech Tips

    FAQs

    1. How do I use qPCR to determine the concentration of my material?
    2. Can I set up my Luna® RT-qPCR at room temperature?
    3. What is the difference between probe- and dye-based versions of the Luna® qPCR Mixes?
    4. Should I use probe- or dye-based detection for my qPCR assays?
    5. How should I design primers for Luna® qPCR?
    6. How long should my amplicon be for qPCR?
    7. Why is the Luna® qPCR Mix blue? Will this dye interfere with detection?
    8. Can I run the Luna® qPCR Mix on my qPCR instrument?
    9. Can I use fast cycling conditions with the Luna® qPCR Mix?
    10. Do I need to add ROX?
    11. How many dilutions should I use to make a standard curve?
    12. Why does NEB recommend 40-45 cycles?
    13. Does the Luna® qPCR Mix contain dUTP? Can I use carryover contamination prevention methods?
    14. Why do I have multiple peaks in my melt curve?
    15. How can I distinguish non-template amplification (NTC) from real products?
    16. Why do I see amplification curves in my NTC samples?
    17. How much primer should I use with the Luna® Universal RT-qPCR Kit?
    18. How do I choose between one-step RT-qPCR and two-step RT-qPCR?
    19. What RNA samples can be used in qPCR with the Luna® Mix?
    20. How much RNA template should I use in my RT-qPCR reaction?
    21. Can I use longer targets in one-step RT-qPCR?
    22. What temperature should I use for cDNA synthesis with Luna® RT-qPCR kits?
    23. Should I include a no Luna® WarmStart® Enyzme Mix control (-RT Control)?
    24. What is the fluorescent, double-stranded DNA binding dye in the Luna® qPCR master mix?
    25. Is the Monarch® Total RNA Miniprep Kit compatible with Luna® RT-qPCR Reagents?
    26. Are the Monarch RNA Cleanup Kits compatible with Luna RT-qPCR reagents?
    27. Can I use shorter cycling times?

    Protocols & Manuals

    Manuals

    The Product Manual includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name these document files: manual[Catalog Number].)

    Protocols

    1. Luna® Universal One-Step RT-qPCR Kit Protocol (E3005)

    Other Tools & Resources

    Featured Articles

    • Development of a high-throughput data analysis method for quantitative real-time PCR (qPCR)
    • Optimization Tips for Luna® One-Step RT-qPCR

    Troubleshooting Guides

    Luna® One-Step RT-qPCR Troubleshooting Guide

    Interactive Tools

    Quality & Safety

    Quality Assurance Statement

    Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

    Specifications

    The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

    Certificate Of Analysis

    The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

    Safety DataSheets

    The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

    Nuclease-free Water
    Luna® WarmStart® RT Enzyme Mix
    Luna® Universal One-Step Reaction Mix