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Fluorescence-based quantitative polymerase chain reaction (qPCR) is the gold standard for the detection and quantification of nucleic acids due to its sensitivity and specificity. Luna products are optimized for qPCR or RT-qPCR, and are available for either intercalating dye or probe-based detection methods. All Luna products provide robust performance on diverse sample sources and target types.

The following Luna qPCR products contain a universal passive reference dye that enables compatibility with a variety of real-time instruments, including those that require No ROX, Low ROX, or High ROX passive reference dye concentrations:

Luna Universal qPCR Master Mix (NEB #M3003)
Luna Universal Probe qPCR Master Mix (NEB #M3004)
Luna Universal One-Step RT-qPCR Kit (NEB #E3005)
Luna Universal Probe One-Step RT-qPCR Kit (NEB #E3006)
Luna Probe One-Step RT-qPCR 4X Mix with UDG (NEB #M3019)
LyoPrime Luna® Probe One-Step RT-qPCR Mix with UDG (NEB #L4001)

For instruments that do not utilize ROX normalization, the Luna Probe One-Step RT-qPCR Kit (No ROX) (NEB #E3007) contains no reference dye. If ROX normalization is desired, ROX can be added; this is only necessary with the E3007 product.

The Luna Probe One-Step RT-qPCR 4X Mix with UDG (NEB #M3019) supports robust, sensitive detection and quantitation of up to 5 targets in a multiplexed reaction. It is supplied at a 4X concentration and enables higher amounts of sample input, which is relevant for applications where RNA present in low abundance is of interest, such as pathogen detection. For simplified reaction setup, the single tube master mix format consolidates components for the one-step RT-qPCR reaction. It also includes dUTP and UDG in the mix for reduced risk of carryover contamination.

The LyoPrime Luna Probe One-Step RT-qPCR Mix with UDG (NEB #L4001) offers the same versatile features and strong performance as the M3019 product in a lyophilized format. For more information on lyophilized products used for RNA detection, please visit www.neb.com/lyoprime.

The Luna SARS-CoV-2 RT-qPCR Multiplex Assay Kit (NEB #E3019) combines the M3019 product with an optimized SARS-CoV-2 Primer/Probe Mix specific to two regions of the SARS-CoV-2 virus N gene [based on sequences provided by the Centers for Disease Control and Prevention (CDC)]. The probes have been modified to contain different fluorophores (N1: HEX; N2: FAM) to enable simultaneous observation on two different channels of a real-time instrument. To ensure the integrity of the input material and absence of inhibition, an internal control (IC) primer and probe set, designed to amplify the human RNase P gene, is also included in the primer mix. The reverse primer of this target has been modified from the CDC design to target an exon/exon boundary to reduce background amplification from possible contaminating genomic DNA. Amplification of the IC is observed in the Cy5 channel. A positive control template (SARS-CoV-2 N gene cloned into a plasmid) is also provided in this kit.

The other Luna products contain dUTP and enable carryover prevention when reactions are treated with Antarctic Thermolabile UDG (NEB #M0372). A blue visible dye assists in tracking the reagents when pipetting into clear, multi-welled PCR plates. The reverse transcriptase featured in the Luna One-Step RT-qPCR products is an engineered WarmStart® enzyme developed for robust performance and increased thermostability. These features, combined with rapid, sensitive and precise real-time qPCR performance, make Luna the universal choice for all your qPCR and RT-qPCR experiments.

For two-step RT-qPCR, the LunaScript® RT SuperMix Kit (NEB #E3010) offers a fast (less than 15 min), robust, and sensitive option for cDNA synthesis upstream of your Luna qPCR experiment. The supermix contains a blue tracking dye allowing you to easily track your samples throughout the RT-qPCR workflow.

The LunaScript RT Master Mix Kit (Primer-free) (NEB #E3025) features an optimized 5X master mix containing all the necessary components for first strand cDNA synthesis, except primers. The mix is compatible with random primers, oligo dT primers, and gene-specific primers, enabling maximum cDNA synthesis flexibility for 2-step RT-qPCR, 2-step RT-PCR, and RNA-seq applications.

The Luna Cell Ready Lysis Module (NEB #E3032) and kits are designed for direct RNA quantitation from cell lysate, bypassing traditional RNA extraction and purification steps. Coordinated cell lysis, RNA release, and genomic DNA removal is achieved in a 15 min protocol, with optimal RT-qPCR analysis obtained when the cell lysate is paired with Luna Universal One-Step RT-qPCR Kits.

Make a simpler choice 

  • Convenient master mix and supermix formats with user-friendly protocols simplify reaction setup
  • Non-interfering, visible tracking dye helps to eliminate pipetting errors
  • Read our white paper on key considerations for optimal lyophilized reagent development
Learn more

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

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Optimize your One-Step RT-qPCR with Luna WarmStart® Reverse Transcriptase 

  • Novel, thermostable reverse transcriptase (RT) improves performance
  • WarmStart RT paired with Hot Start Taq. Unique aptamer technology increases reaction specificity and robustness.
  • Skip RNA purification and go direct from cells to RT-qPCR analysis with Luna Cell Ready Module and kits

Learn more

Luna qPCR and RT-qPCRInterested in learning more about
Luna qPCR & RT-qPCR products?
Download our product brochure

 

 

 

Available products:

Luna Universal qPCR Master Mix

Rapid, sensitive, and precise dye-based qPCR detection and quantitation of target DNA and cDNA sequences 
Luna Universal Probe qPCR Master Mix

Rapid, sensitive, and precise probe-based qPCR detection and quantitation of target DNA and cDNA sequences
Luna Universal One-Step RT-qPCR Kit

Includes everything you need for rapid, sensitive, and precise dye-based qPCR detection and quantitation of RNA targets
Luna Universal Probe One-Step RT-qPCR Kit

Includes everything you need for rapid, sensitive, and precise probe-based qPCR detection and quantitation of RNA targets
Luna Probe One-Step RT-qPCR 4X Mix with UDG

4X mix enables sensitive detection of target RNA sequences, with robust performance in multiplex applications of up to 5 targets
LyoPrime Luna Probe One-Step RT-qPCR Mix with UDG

Lyophilized format enables sensitive detection of target RNA sequences with room temperature stability
Luna SARS-CoV-2 RT-qPCR Multiplex Assay Kit

For real-time qualitative detection of SARS-CoV-2 nucleic acid using hydrolysis probes
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)

Single-tube 5X master mix contains Luna Reverse Transcriptase, dNTPs, and Murine RNase Inhibitor
Luna Cell Ready One-Step RT-qPCR Kit

All the necessary components for direct, dye-based RNA detection and quantitation from cell lysate, bypassing the need for RNA extraction and purification
Luna Cell Ready Probe One-Step RT-qPCR Kit

All the necessary components for direct, probe-based RNA detection and quantitation from cell lysate, bypassing the need for RNA extraction and purification
Luna Cell Ready Lysis Module

Coordinates cell lysis, RNA release, and genomic DNA removal simultaneously, resulting in cell lysate ready for direct RT-qPCR analysis with Luna Universal One-Step RT-qPCR kits

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Videos

  • overview of qPCR

    Overview of qPCR

    Learn the basics of qPCR in this short animation.                                                                                                  

  • NEBTv_Ep16_Thumb

    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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