1 1

New England Biolabs

NEB LunaScriptRT SuperMix Kit (Cat No. E3010)

NEB LunaScriptRT SuperMix Kit (Cat No. E3010)

Request quotation at the end of the product description.

LunaScript RT SuperMix Kit (NEB #E3010) includes supermix, No-RT control mix, and nuclease-free water.

Supermix only option now available, please see LunaScript RT SuperMix (NEB #M3010).

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

 

  • 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 include nuclease-free water and No-RT Control Mix

 

Catalog # Concentration Size
E3010S Not Applicable 25 reactions
E3010L Not Applicable 100 reactions

 

 

 

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 ΔCqmeasures low input detection and lack of non-template control (NTC) amplification (ΔCq = average Cq of NTC - average Cqof 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.




 

 

Product Name: NEB LunaScriptRT SuperMix Kit (Cat No. E3010)

View full details

-----

Instant Quote Request

Click Here For Instructions

While we cannot display all our products on this site, this tool provides you with access to our entire product lineup. Simplify your quote requests by using this platform. Begin exploring today!

 

Step1: Click "Search Products"
Browse thru our extensive list of products and add them to your request by clicking the "+" button on each product entry. 

Step 2: Click "Review Requested Items"
Review the list of your requested items. You may also add more items by clicking the "+" button at the bottom right.

Step 3: Click "Proceed Quote Request"
Finalize the request by inputting your details so we know where to send the quotation.

And that's it! 

Please click the "Accept" button below to proceed.

.