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LunaScript® RT SuperMix

Catalog # Concentration Size List Price Quantity Your Price
M3010E 5 X 2500 reactions $11,061.00
$9,954.90
M3010L 5 X 100 reactions $638.00
$574.20
M3010X 5 X 500 reactions $2,553.00
$2,297.70
Catalog # Size List Price Your Price
M3010E 2500 reactions $11,061.00
$9,954.90
M3010L 100 reactions $638.00
$574.20
M3010X 500 reactions $2,553.00
$2,297.70
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
  • Single-tube supermix contains random hexamer and oligo-dT primers, dNTPs, Murine RNase Inhibitor, and Luna® Reverse Transcriptase
  • Less than 15 minute first-strand cDNA synthesis protocol
  • Novel, thermostable Luna Reverse Transcriptase improves performance 
  • Combine with Luna qPCR master mixes (NEB #M3003, M3004) for robust RT-qPCR results
  • Eliminate pipetting errors with non-interfering, visible tracking dye
  • Primer-free version (NEB #E3025) available
  • Does not include nuclease-free water or a No-RT Control Mix

1. Josh Quick 2020. nCoV-2019 sequencing protocol v3 (LoCost). protocols.io 

Featured Videos
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  • Monarch_MiniprepProtocol_VideoThumb
    Avoiding RNase Contamination
  • Monarch_MiniprepProtocol_VideoThumb
  • Monarch_MiniprepProtocol_VideoThumb
    Overview of qPCR
  • Monarch_MiniprepProtocol_VideoThumb
    NEB® TV Ep. 16 – Overview and Applications of qPCR

LunaScript RT SuperMix is an optimized master mix for first strand cDNA synthesis and can be used in amplicon sequencing or 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. LunaScript RT SuperMix contains random hexamer and poly-dT primers, allowing even coverage across the length of the RNA targets. In addition, LunaScript RT SuperMix contains a blue dye, providing a visual indicator that can be followed throughout the two-step RT-qPCR process. The product offers robust, linear, and sensitive detection using total RNA inputs as high as 1 µg and as low as single copies of RNA.

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

In comparison to the LunaScript kit version (NEB #E3010), this LunaScript product does not contain a No-RT Control Mix or nuclease-free water. More details can be found below:

Figure 1: Comparison of LunaScript Products





Figure 2: Product options for one-step and two-step RT-qPCR workflows





Figure 3: LunaScript RT SuperMix offers exceptional sensitivity, linearity and reproducibility in two-step RT-qPCR workflows



RNA 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 was then quantitated by qPCR using 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.




Figure 4: LunaScript RT SuperMix supports even coverage of long transcripts



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 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.




Figure 5: LunaScript RT SuperMix demonstrates superior linear detection of RNA targets



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 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.




Figure 6: At just 13 minutes, LunaScript RT SuperMix offers the shortest available first-strand cDNA synthesis protocol 



Comparison of recommended protocols for cDNA synthesis. LunaScript RT SuperMix requires the shortest reaction time and tolerates elevated temperatures, reducing complications from RNA secondary structure.




Figure 7: LunaScript RT SuperMix exhibits exceptional stability at room temperature



LunaScript RT SuperMix and other commercial products were left at 25°C for 15 days, and then 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 Luna Universal qPCR Master Mix (NEB #M3003). Results were evaluated for efficiency and ΔCq, where ΔCq measures low input detection and lack of no-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.




Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  LunaScript® RT SuperMix M3010LVIAL -20 1 x 0.4 ml 5 X
  LunaScript® RT SuperMix M3010LVIAL -20 5 x 0.4 ml 5 X
  LunaScript® RT SuperMix M3010EVIAL -20 1 x 10 ml 5 X
LunaScript® RT SuperMix | New England Biolabs

LunaScript® RT SuperMix

Supermix only version; this product does not include a No-RT control mix or nuclease-free water.

LunaScript RT SuperMix Kit (NEB https://www.neb.com/en/#E3010) is a kit that includes supermix, No-RT control mix, and nuclease-free water.

Featured in the ARTIC SARS-CoV-2 sequencing workflow (1) and a component in the NEBNext® ARTIC SARS-CoV-2 kits.


Catalog https://www.neb.com/en/#M3010

  • Single-tube supermix contains random hexamer and oligo-dT primers, dNTPs, Murine RNase Inhibitor, and Luna® Reverse Transcriptase
  • Less than 15 minute first-strand cDNA synthesis protocol
  • Novel, thermostable Luna Reverse Transcriptase improves performance 
  • Combine with Luna qPCR master mixes (NEB https://www.neb.com/en/#M3003, M3004) for robust RT-qPCR results
  • Eliminate pipetting errors with non-interfering, visible tracking dye
  • Primer-free version (NEB https://www.neb.com/en/#E3025) available
  • Does not include nuclease-free water or a No-RT Control Mix

1. Josh Quick 2020. nCoV-2019 sequencing protocol v3 (LoCost). protocols.io 

Featured Video

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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.
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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.The buyer/user has a non-exclusive license to use the vector for Research Purposes Only. Commercial use of this vector requires a license from New England Biolabs, Inc.

U.S. Patent No. 5,643,758

Appln. No. WO2007/120809

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