LunaScript® RT SuperMix Kit

Catalog #	Concentration	Size	List Price	Quantity	Your Price
E3010L		100 reactions	$711.00		$639.90
E3010S		25 reactions	$232.00		$208.80

Catalog #	Size	List Price	Your Price
E3010L	100 reactions	$711.00	$639.90
E3010S	25 reactions	$232.00	$208.80

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:

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.*

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.

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 ΔC_q measures low input detection and lack of non-template control (NTC) amplification (ΔC_q = average C_q of NTC - average C_q of lowest input). Green box indicates target performance criteria (Efficiency = 90-110%, ΔC_q ≥ 3).

Learn more about this “Dots in Boxes” visualization method.

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.

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 ΔC_q, where ΔC_q measures low input detection and lack of no-template control (NTC) amplification (ΔC_q = average C_q of NTC - average Cq of lowest input). Green box indicates target performance criteria (Efficiency = 90-110%, ΔC_q ≥ 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

E3010S -20

LunaScript^® RT SuperMix	M3010SVIAL	-20	1 x 0.1 ml	5 X
No-RT Control Mix	M3011SVIAL	-20	1 x 0.1 ml	5 X
Nuclease-free Water	B1502AVIAL	-20	1 x 1.5 ml

E3010L -20

LunaScript^® RT SuperMix	M3010LVIAL	-20	1 x 0.4 ml	5 X
No-RT Control Mix	M3011LVIAL	-20	1 x 0.4 ml	5 X
Nuclease-free Water	B1502AVIAL	-20	4 x 1.5 ml

Properties & Usage

Companion Products

Protocols

DataCards

manualE3010

FAQs

Citations

Quality Control Assay

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.

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 Data Sheets

Legal And Disclaimer

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.