Luna® Universal qPCR & RT-qPCR
Optimize One-Step RT-qPCR with unique WarmStart® technology

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Lighting the way – Luna® Universal qPCR & RT-qPCR banner with luna moth

The Luna One-Step RT-qPCR kits contain a novel, in silico-designed reverse transcriptase (RT) engineered for improved performance. Both the Luna WarmStart Reverse Transcriptase and Hot Start Taq DNA Polymerase, included in these kits, utilize a temperature-sensitive, reversible aptamer, which inhibits activity below 45°C. This enables room temperature reaction setup and prevents undesired non-specific activity. Furthermore, the WarmStart RT has increased thermostability, improving performance at higher reaction temperatures.

Improve performance with our novel, thermostable reverse transcriptase

  • Robust RT delivers improved specificity, sensitivity and efficiency
  • Increased thermostability adds flexibility to reaction setup and improved specificity
Figure 1: Luna WarmStart Reverse Transcriptase exhibits exceptional sensitivity, reproducibility and RT-qPCR performance

Amplification plot, standard curve and melt curve showing sensitivity and reproducibility of performance
RT-qPCR targeting human GAPDH was performed using the Luna Universal One-Step RT-qPCR Kit, featuring Luna WarmStart Reverse Transcriptase, over an 8-log range of input template concentrations (0.1 pg – 1 μg Jurkat total RNA) with 8 replicates at each concentration. NTC = non-template control.

Achieve better control and better results: WarmStart Luna RT paired with Hot Start Taq

  • WarmStart/Hot Start technology ensures tight control and prevents spurious amplification
  • Unique WarmStart technology is not available with other non-NEB commercially-available products
Figure 2: WarmStart Luna RT prevents spurious amplification resulting from room-temperature pre-incubation

Amplfication plots and melt curves of Luna® Universal One-Step RT-qPCR Kit as compared to ABI SYBR® Green 1-Step Kit
RT-qPCR targeting human ribosomal protein L32 was performed before and after a 24-hour incubation at room temperature, with triplicate reactions for a 5-log range of input human (Jurkat) total RNA and a non-template control. The Luna Universal One-Step RT-qPCR Kit featuring Luna WarmStart Reverse Transcriptase exhibited robust performance and no detectible non-template amplification, either with or without a 24 hour 25°C pre-incubation, while the ABI 1-Step Kit, featuring a non-WarmStart reverse transcriptase, exhibited significant non-template amplification.

Greater thermostability means greater versatility

  • Increased RT reaction temperature is ideal for difficult targets
Figure 3: The increased thermostability of WarmStart Luna Reverse Transcriptase improves performance at increased reaction temperatures

Graphs demonstrating thermostability of Luna WarmStart Reverse Transcriptase as compared to MMLV Reverse Transcriptase
RT-qPCR experiments targeting human ribosomal protein L32 RNA were performed in triplicate over a 5-log range of input human (Jurkat) total RNA (5 pg – 50 ng) using an initial 10 min RT step performed at 50°C – 60°C, as indicated.

A. Luna WarmStart Reverse Transcriptase (recommended incubation temperature: 55°C) exhibited rapid Cq values (bar graph) and robust RT-qPCR performance (amplification plots) at each temperature, indicating that efficient reverse transcription was not perturbed by reaction temperature alterations.

B. In contrast, a commercially available MMLV (recommended incubation temperature: 50°C) exhibited delayed (increased) Cq values, poorer performance, and loss of low-input detection at elevated temperatures, consistent with loss of RT activity.



Available products:

Luna Universal qPCR Master Mix

Dye-based qPCR for DNA and cDNA
Luna Universal Probe qPCR Master Mix

Probe-based qPCR for DNA and cDNA
Luna Universal One-Step RT-qPCR Kit

Dye-based one-step RT-qPCR for RNA
Luna Universal Probe One-Step RT-qPCR Kit

Probe-based one-step RT-qPCR for RNA
Luna Probe One-Step RT-qPCR 4X Mix with UDG

4X mix for probe-based one-step RT-qPCR
LyoPrime Luna Probe One-Step RT-qPCR Mix with UDG

Single tube with blue tracking dye includes everything needed for first strand cDNA synthesis in two-step RT-qPCR workflows

Luna Direct Probe One-Step RT-qPCR 4X Mix with UDG

Supports direct probe-based one-step RT-qPCR from crude or purified RNA samples 

Luna Direct Probe qPCR 4X Master Mix with UDG

Supports direct probe-based qPCR from crude or purified DNA samples 
Luna SARS-CoV-2 RT-qPCR Multiplex Assay Kit

Flexible first-strand cDNA synthesis with user-selected primers
LunaScript RT SuperMix Kit

Single tube with blue tracking dye includes everything needed for first strand cDNA synthesis in two-step RT-qPCR workflows
LunaScript RT Master Mix Kit (Primer-free)

Flexible first-strand cDNA synthesis with user-selected primers
Luna Cell Ready One-Step RT-qPCR Kit

Cell lysis module + dye-based RT-qPCR kit for direct detection from cell lysates
Luna Cell Ready Probe One-Step RT-qPCR Kit

Cell lysis module + probe-based RT-qPCR kit for direct detection from cell lysates
Luna Cell Ready Lysis Module

Cell lysis module alone

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Videos

  • Overview of qPCR

    Learn the basics of qPCR in this short animation.                                                                                                  

  • NEB® TV Ep. 16 – Overview and Applications of qPCR

    Learn the basics of qPCR, and hear more about our Luna products, including our novel method for data analysis.

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SYBR® is a registered trademark of Molecular Probes, Inc.
ARRAYSCRIPT™ is a trademark of Thermo Fisher Scientific,Inc.