Product Class: Other

Luna® Universal qPCR Master Mix

Product Introduction

Rapid, sensitive and precise dye-based qPCR detection and quantitation of target DNA and cDNA sequences.

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
Catalog # Size Concentration
M3003S 200.0 reactions 2 X
M3003L 500.0 reactions 2 X
M3003X 1000.0 reactions 2 X
M3003E 2500.0 reactions 2 X

Product Information

Description

Dye-based quantitative PCR (qPCR) uses real-time fluorescence of a double-stranded DNA (dsDNA) binding dye, most commonly SYBR® Green I, to measure DNA amplification during each cycle of a PCR. 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.

The Luna Universal qPCR Master Mix is an optimized 2X reaction mix for real-time qPCR detection and quantitation of target DNA sequences using the SYBR®/FAM channel of most real-time qPCR instruments. It contains Hot Start Taq DNA Polymerase and has been 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). It also features dUTP for carryover prevention and a non-fluorescent, visible dye to monitor reaction setup. This dye does not spectrally overlap with fluorescent dyes used for qPCR and will not interfere with real-time detection.

The master mix formulation is supplied at 2X concentration and contains all PCR components required for amplification and quantitation of DNA except primers and DNA template. Genomic DNA or cDNA of interest can be quantitated with Luna qPCR, and existing as well as commercial qPCR assay primer sequences can be used.


Figure 1: The Luna Universal qPCR Master Mix offers exceptional sensitivity, reproducibility and qPCR performance
qPCR targeting human GAPDH was performed using the Luna Universal qPCR Master Mix over a 6-log range of input template concentrations (20 ng – 0.2 pg Jurkat-derived cDNA) with 8 replicates at each concentration. cDNA was generated from Jurkat total RNA using the NEB Protoscript® II First Strand cDNA Synthesis Kit (NEB #E6560).


Figure 2: The Luna Universal qPCR Master Mix provides sensitive and accurate detection and quantitation across a wide variety of DNA sources
Luna
The Luna Universal qPCR Master Mix is compatible with a broad range of genomic DNA sources. qPCR targets were quantitated with 50 ng – 5 pg genomic DNA as input using an ABI 7500 Fast real-time instrument. Genomic DNA was purified by typical column-based methods. In these examples, strong performance can be observed in the amplification of ACTB (encoding β-actin) from Mouse kidney genomic DNA, psbB (Photosystem II CP47 reaction center protein PsbB) from Tobacco, and RDN18 (18S ribosomal RNA) from Yeast.


Figure 3: Extensive performance evaluation of commercially available dye-based qPCR reagents demonstrates the robustness and specificity of Luna
Luna
qPCR reagents from NEB and other manufacturers were tested across 16–18 qPCR targets varying in abundance, length and %GC, using either Jurkat genomic DNA or Jurkat-derived cDNA as input (10 genomic DNA targets and 8 cDNA targets on Bio-Rad real-time instrument, 9 genomic and 7 cDNA targets on ABI instrument). For each testing condition, data was collected by 2 users and according to manufacturer’s specifications. 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 major manufacturers are shown: Bio-Rad, SsoAdvanced™ Universal SYBR® Green Supermix; Roche, FastStart™ SYBR Green Master; QIAGEN, QuantiTect® SYBR Green PCR Kit; ABI, PowerUP™ SYBR Green Master Mix; Promega®, GoTaq® qPCR Master Mix. The Luna Universal qPCR Master Mix outperformed all other reagents tested.

Learn more about our comprehensive qPCR/RT-qPCR testing and “dots in boxes” data visualization

 

Figure 4: The Luna Universal qPCR Master Mix contains an inert blue tracking dye to eliminate pipetting errors.


 


This product is related to the following categories:
Luna® qPCR & RT-qPCR Products,
PCR, qPCR & Amplification Technologies Products,
This product can be used in the following applications:
qPCR & RT-qPCR,
DNA Amplification, PCR & qPCR

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

Properties & Usage

Protocols, Manuals & Usage

Protocols

  1. Luna® Universal qPCR Master Mix Protocol (M3003)
  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) 

Manuals

The Product Manual includes details for how to use the product, as well as details of its formulation and quality controls.

Usage & Guidelines

Application Notes

Tools & Resources

Web Tools

FAQs & Troubleshooting

FAQs

  1. How do I use qPCR to determine the concentration of my material?
  2. Can I set up my Luna® 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 instrument settings 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. What samples can be used in qPCR with the Luna® Mix?
  18. Can I use cDNA? Does it matter how I make it?
  19. How much template material can I use in Luna® qPCR?
  20. How much primer should I use for the Luna® Universal qPCR Master Mix?
  21. Can I use shorter cycling times?
  22. What is the fluorescent, double-stranded DNA binding dye in the Luna® qPCR master mix?

Troubleshooting

Quality, Safety & Legal

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.

Specification Change Notifications

Effective May 2, 2022, a secondary bottle supplier has been qualified. The alternative 30 mL bottle has a different cap color and may be used for relevant pack sizes with no change in product performance or stability.

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.

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

Licenses

Nucleic acid-based aptamers for use with thermophilic DNA polymerases are licensed exclusively by New England Biolabs, Inc. from SomaLogic, Inc. New England Biolabs, Inc. gives the Buyer/User a non-exclusive license to use the Luna Universal qPCR Master Mix for Research Uses Only (RUO). Commercial use of this product may require a license from New England Biolabs, Inc. For additional information or to inquire about commercial use, please contact busdev@neb.com. 

Trademarks

BIO-RAD® is a registered trademark of Bio-Rad Laboratories.
FASTSTART™ is a trademark of Roche.
GOTAQ® and PROMEGA® are registered trademarks of Promega Corporation.
POWERUP™ is a trademark of Thermo Fisher Scientific.
QIAGEN® and QUANTITECT® are registered trademarks of Qiagen, Inc.
QUANTSTUDIO® is a registered trademark of Life Technologies Corporation.
SOMAMER® is a registered trademark of Somalogic, Inc.
SSOADVANCED™ is a trademark of Bio-Rad Laboratories.
STEPONEPLUS® is a registered trademark of Applied Biosystems, LLC.
SYBR® is a registered trademark of Molecular Probes, Inc.

Other Products You May Be Interested In