Item# Description List Price Web Price Qty
    E3010L LunaScript SuperMix Kit - 100 rxns $588.00
    $529.20
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    E3010S LunaScript SuperMix Kit - 25 rxns $189.00
    $170.10
    ADD TO CART
    *On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

    Product Introduction

    The LunaScript™ RT SuperMix Kit is optimized for cDNA synthesis in a two-step RT-qPCR workflow.

    Make a simpler choice
    • Single-tube SuperMix contains random hexamer and oligo-dT primers, dNTPs, Murine RNase Inhibitor, and Luna® Reverse Transcriptase 
    • Non-interfering, visible tracking dye helps to eliminate pipetting errors
    • Combine with Luna qPCR master mixes for robust RT-qPCR results
    Experience best-in-class performance
    • Less than 15 minute first-strand cDNA synthesis protocol
    • All Luna products have undergone rigorous testing to optimize specificity, sensitivity, accuracy and reproducibility
    • A comprehensive evaluation of commercially-available qPCR and RT-qPCR reagents demonstrates superior performance of Luna products
    Illuminate your two-step RT-qPCR
    • Novel, thermostable Luna Reverse Transcriptase improves performance 
    • Consistent linearity, sensitivity, and capacity for reliable RNA quantification

    Product Information

    Description

    LunaScript_Sample_Banner

    LunaScript RT SuperMix is an optimized master mix containing all the necessary components for first strand cDNA synthesis in the context of a two-step RT-qPCR workflow. It features the thermostable Luna Reverse Transcriptase, which supports cDNA synthesis at elevated temperatures. Murine RNase Inhibitor is also included to protect template RNA from degradation. The LunaScript RT SuperMix contains random hexamer and poly-dT primers, allowing even coverage across the length of the RNA targets. In addition, the LunaScript RT SuperMix contains a blue dye, providing a visual indicator that can be followed throughout the two-step RT-qPCR process. The LunaScript RT SuperMix offers robust, linear, and sensitive detection using total RNA inputs as high as 1 µg and as low as single copies of RNA.

    Comparison of one-step and two-step RT-qPCR workflows
    Fig1_LunaScript_OneStep_TwoStep_RTqPCR_0118
    The LunaScript RT SuperMix Kit offers exceptional sensitivity, linearity and reproducibility in two-step RT-qPCR workflows
    Fig2_LunaScript_RTqPCR_0118
    RNA was converted to cDNA using the 1X LunaScript RT SuperMix in 20 μl reactions using standard reaction conditions (25°C/2 min, 55°C/10 min, 95°C/1 min). cDNA was then quantitated by qPCR using the Luna Universal qPCR Master Mix (NEB #M3003) and 1 μl of cDNA product as template, with triplicate reactions at each input concentration. A. A serial dilution of Jurkat total RNA (1 μg – 1 pg) was converted to cDNA and then quantitated by qPCR using a β-actin target. B. ERCC (External RNA Controls Consortium) mix1 RNA containing 5x109 to 50 copies of ERCC00130 (~10 ng – 10 fg) was converted to cDNA and then quantitated by qPCR.
    The LunaScript RT SuperMix Kit supports even coverage of long transcripts
    Fig3_LunaScript_LongTranscriptCoverage_0118
    Eight pairs of primers were designed to cover the entire 15.2 kb HERC1 transcript. Jurkat total RNA (1 μg - 1 ng) was converted to cDNA using 1X LunaScript RT SuperMix in 20 μl reactions using standard reaction conditions (25°C/2 min, 55°C/10 min, 95°C/1 min). cDNA coverage was then evaluated by qPCR using the Luna Universal qPCR Master Mix (NEB #M3003) and 1 μl of cDNA product as template, with duplicate reactions for each target at each input concentration.
    The LunaScript RT SuperMix Kit demonstrates superior linear detection of RNA targets
    Fig4_LunaScript_LinearDetection_0118
    Commercially available cDNA supermixes were used according to manufacturer’s recommendations to generate cDNA from 1 μg – 100 pg human (Jurkat) total RNA. cDNA products were then evaluated by qPCR using eight targets varying in abundance, length and %GC. qPCR detection was performed using the Luna Universal qPCR Master Mix (NEB #M3003) or Luna Universal Probe qPCR Master Mix (NEB #M3004). Results were evaluated for efficiency and ΔCq, where ΔCq measures low input detection and lack of non-template control (NTC) amplification (ΔCq = average Cq of NTC - average Cq of lowest input). Green box indicates target performance criteria (Efficiency = 90-110%, ΔCq ≥ 3).

    Learn more about this “Dots in Boxes” visualization method.
    At just 13 minutes, the LunaScript RT SuperMix Kit offers the shortest available first-strand cDNA synthesis protocol
    Fig5_LunaScript_CyclingConditions_0118
    Comparison of recommended protocols for cDNA synthesis. The LunaScript RT SuperMix Kit requires the shortest reaction time and tolerates elevated temperatures, reducing complications from RNA secondary structure.

    Kit Components

    The following reagents are supplied with this product:

    Store at (°C) Concentration
    Nuclease-free Water -20
    LunaScript RT SuperMix -20 5 X
    No-RT Control Mix -20 5 X
    Product Categories:
    Luna® qPCR & RT-qPCR Products
    Applications:
    Reverse Transcription (cDNA Synthesis)

    Properties and Usage

    Storage Temperature

    -20°C

    Faqs & Tech Tips

    FAQs

    1. Can I use  RNA samples purified from different storage conditions such as RNALater in my LunaScript®  cDNA synthesis reactions?
    2. Can I add gene-specific primers in my LunaScript®  cDNA synthesis reaction?
    3. Can I set up my cDNA synthesis reaction at room temperature when using the LunaScript®  RT SuperMix?
    4. Can I store the LunaScript®  RT SuperMix at room temperature or 4°C?
    5. Can I use a shorter incubation time for my cDNA synthesis reaction when using the LunaScript®  RT SuperMix?
    6. Can I use LunaScript® cDNA products directly in qPCR? How much can I use for detection in a qPCR reaction?
    7. How do I choose between LunaScript® RT SuperMix Kit and ProtoScript first strand cDNA synthesis kits?
    8. How do I know whether my template RNA is of good quality?
    9. How much RNA template should I use in my LunaScript®  cDNA synthesis reactions?
    10. How much cDNA product can I use for detection in a qPCR reaction?
    11. How should I store LunaScript®  cDNA products?
    12. How should I choose between a one-step RT-qPCR or two-step RT-qPCR approach?
    13. Should I include a No-Reverse Transcriptase (RT) control reaction when setting up my cDNA synthesis reaction?
    14. What qPCR reagents do you recommend for detection of cDNA products?
    15. What RNA samples can be used in the LunaScript®  cDNA synthesis reactions?
    16. What temperature should I use for my LunaScript®  cDNA synthesis reaction?
    17. Why do I have low cDNA yields?
    18. Will the blue dye in the LunaScript®  RT SuperMix interfere with detection?
    19. Are the Monarch RNA Cleanup Kits compatible with Luna RT-qPCR reagents?

    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. Protocol for LunaScript®  RT SuperMix Kit (E3010)
    2. Protocol for Two-step RT-qPCR using the LunaScript™ RT SuperMix Kit (NEB #E3010) and the Luna® Universal qPCR Master Mix (NEB #M3003) or Luna Universal Probe qPCR Master Mix (NEB #M3004) 

    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.