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NOW INCLUDES THE LUNASCRIPT RT SUPERMIX KIT Luna ® Universal qPCR & RT-qPCR LIGHTING THE WAY be INSPIRED drive DISCOVERY stay GENUINE

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Page 1: Luna Universal qPCR & RT-qPCR - New England Biolabs GmbH€¦ · Luna ® Universal qPCR & RT-qPCR LIGHTING THE WAY ™ beINSPIRED driveDISCOVERY stayGENUINE. 2 Fluorescence-based

NOW INCLUDES THE LUNASCRIPT™ RT SUPERMIX KIT

Luna® Universal qPCR & RT-qPCR LIGHTING THE WAY™

be INSPIRED drive DISCOVERY stay GENUINE

Page 2: Luna Universal qPCR & RT-qPCR - New England Biolabs GmbH€¦ · Luna ® Universal qPCR & RT-qPCR LIGHTING THE WAY ™ beINSPIRED driveDISCOVERY stayGENUINE. 2 Fluorescence-based

2

Fluorescence-based quantitative real-time qPCR (qPCR) is the “gold standard” for the detection and quantitation of nucleic acids due to its sensitivity and specificity. Luna products from NEB® 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.

Each Hot Start Taq-based Luna qPCR master mix has been formulated with a unique passive reference dye that is compatible across a wide variety of instrument platforms, including those that require a ROX reference signal. This means that no additional components are required to ensure machine compatibility. The mixes also contain dUTP, enabling carryover prevention when reactions are treated with NEB’s Antarctic Thermolabile UDG (NEB #M0372). A blue visible dye assists in tracking the reagents when pipetting into clear, multi-welled PCR plates. These features, combined with rapid, sensitive and precise real-time qPCR performance, make Luna the universal choice for all your qPCR experiments.

For two-step RT-qPCR, the LunaScript™ RT SuperMix Kit 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.

Lighting the way Make a simpler choice• One product per application

simplifies selection

• Convenient master mix and supermix formats with 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

Optimize your RT-qPCR:Luna WarmStart® Reverse Transcriptase or LunaScript™ RT SuperMix Kit

• Employs novel, thermostable Reverse Transcriptases (RT) for fast protocols at elevated temperatures, exceptional robustness and sensitivity

• One-Step Kits: the unique WarmStart RT paired with Hot Start Taq increases reaction specificity and robustness

• Two-Step: the convenient LunaScript RT SuperMix Kit optimized for best-in-class two-step protocols includes dNTPs, primers and RNase Inhibitor

Find the right Luna product for your application

Reversetranscription qPCR

Reversetranscription & qPCR

DNAcDNA cDNARNA

DNARNA cDNAOne-StepRT-qPCR

Two-StepRT-qPCR

Luna® Universal One-Step RT-qPCR Kit (NEB #E3005)

Luna Universal Probe One-Step RT-qPCR Kit (NEB #E3006)

LunaScript™ RT SuperMix Kit

(NEB #E3010)

Luna Universal qPCR Master Mix

(NEB #M3003)

Luna Universal Probe qPCR Master Mix

(NEB #M3004)

Visible tracking dye

Reversetranscription qPCR

Reversetranscription & qPCR

DNAcDNA cDNARNA

DNARNA cDNAOne-StepRT-qPCR

Two-StepRT-qPCR

Luna® Universal One-Step RT-qPCR Kit (NEB #E3005)

Luna Universal Probe One-Step RT-qPCR Kit (NEB #E3006)

LunaScript™ RT SuperMix Kit

(NEB #E3010)

Luna Universal qPCR Master Mix

(NEB #M3003)

Luna Universal Probe qPCR Master Mix

(NEB #M3004)

Page 3: Luna Universal qPCR & RT-qPCR - New England Biolabs GmbH€¦ · Luna ® Universal qPCR & RT-qPCR LIGHTING THE WAY ™ beINSPIRED driveDISCOVERY stayGENUINE. 2 Fluorescence-based

3

We tested plates and plates of reactions so you don’t have to

NEB has developed a method to better evaluate the large amount of qPCR data generated in an experiment. The output of this analysis is known as a dot plot, and captures the key features of a successful, high-quality qPCR experiment as a single point. This method of analysis allows many targets and conditions to be compared in a single graph.

For each experiment, triplicate reactions are set up across a five-log range of input template concentrations (Amplification plot, bottom-left). Three non-template control (NTC) reactions are also included, for a total of 18 reactions per condition/target. Efficiency (%) is calculated (Standard plot, top-left) and is plotted against ΔC

q

(dot plot, top-center), which is the difference between the average C

q of the NTC and the lowest input. This parameter captures both

detection of the lowest input and non-template amplification.

Acceptable performance criteria are defined as an Efficiency of 90–110% and a ΔC

q of ≥ 3 (green box – pass).

Other performance criteria are captured using a 5-point quality score (Quality score metrics, top-right). Included are:

1. Linearity of amplification, as indicated by the R2 standard curve

2. Reproducibility, as indicated by the consistency of triplicate C

q values for each input concentration

3. Fluorescence consistency, as indicated by similar endpoint fluorescence (RFU

max)

4. Curve steepness

5. Sigmoid curve shape

Evaluating qPCR results: capturing performance as “dots in boxes”

Breaking it down: how we translate qPCR data into "dots in boxes"

Effi

cien

cy (

%)

∆Cq

-5 0 10 2060

80

100

120

140

Fail

Pass

Qualityscore

012345

Pas

s

qPCR performance dot plot Quality score metrics1. R2 (standard curve)2. Cq reproducibility3. Fluorescence consistency (RFUmax)4. Curve steepness5. Curve shapeR

FU

Cycle

RF

U

Cycle

Pass: quality score 5

Fail: quality score 1

Quality scoreStandard curve

Cq

Log Starting Quantity (pg)

Efficiency = 95.1%

R2 = 0.999

Log Starting Quantity (pg)

Amplification plot

= Cq (NTC) – Cq (5 pg)

Cycle

RF

U

∆Cq

Triplicate standards5 pg – 50 ng total RNA

50 ng

5 ng

0.5 ng

50 pg

5 pg

NTC

Quality Score is represented by the size and fill of the plotted dot, with experiments that pass all performance criteria represented by a solid dot within the box. These scoring methods were built upon the MIQE qPCR/RT-qPCR guidelines (Bustin, S.A. et al. (2009) Clin. Chem. 55, 611-22 and Trombley Hall, A. et al. (2013) PLoS One 8(9):e73845).

Visit LUNAqPCR.de (in Germany & Austria) or LUNAqPCR.fr (in France) for a video explanation of our “dots in boxes” evaluation and to request a sample.

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Experience best-in-class performance for your qPCR & RT-qPCRAll NEB products undergo rigorous testing to ensure optimal performance, and Luna is no exception. We took into consideration numerous important traits when evaluating qPCR, including specificity, sensitivity, accuracy and reproducibility, to develop best-in-class qPCR reagents. Furthermore, we did a comprehensive evaluation of commercially-available qPCR and RT-qPCR reagents, and developed a method of analysis that allows you to quickly compare and evaluate the performance of these products. We wanted to be sure that Luna products will perform to your expectations for all your targets.

Evaluation of commercially-available dye-based qPCR reagents demonstrates the robustness and specificity of 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 a Bio-Rad real-time instrument, 9 genomic and 7 cDNA targets on an 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 lowest input – average Cq of non-template control). 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. NEB’s Luna Universal qPCR Master Mix outperformed all other reagents tested.

Luna products offer exceptional sensitivity, reproducibility and qPCR performance

Quantity (pg)

Cq

Efficiency = 96.6%

R2 = 0.999

34

28

26

24

32

30

22

20

0.1 1 10 102 103 104 105

36

20 ng

2 ng

0.2 ng

20 pg

2 pg

0.2 pg

NTC

Cycle2 383634323028262422201810864 40161412 42 44

1

0.1

n = 8

∆R

n

Amplification plot

Standard curve

Input

Quantity (pg)

Cq

Efficiency = 96.6%

R2 = 0.999

34

28

26

24

32

30

22

20

0.1 1 10 102 103 104 105

36

20 ng

2 ng

0.2 ng

20 pg

2 pg

0.2 pg

NTC

Cycle2 383634323028262422201810864 40161412 42 44

1

0.1

n = 8

∆R

n

Amplification plot

Standard curve

Input

qPCR targeting human GAPDH was performed using the Luna Universal Probe 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). NTC = non-template control

86%

NEB

67%

Bio-Rad®

61%

Roche

47%

QIAGEN®

47%

ABI

14%

Promega®

78%

NEB

47%

ABI

80

60

40

20

0

100

80

60

40

20

0

100

70

80

90

100

110

120

130 130

0 10 20 0 10 20 0 10 20 0 10 20 0 10 20 0 10 20 0 10 20 0 10 2070

80

90

100

110

120

QualityScore

543210

pass

Bio-Rad real-timeinstrument

Tar

gets

pas

sing

perf

orm

ance

crit

eria

(%

)E

ffici

ency

(%

)

ABI real-timeinstrument

ΔCq ΔCq

Page 5: Luna Universal qPCR & RT-qPCR - New England Biolabs GmbH€¦ · Luna ® Universal qPCR & RT-qPCR LIGHTING THE WAY ™ beINSPIRED driveDISCOVERY stayGENUINE. 2 Fluorescence-based

5

Experience best-in-class performance for your qPCR & RT-qPCRTobacco leaf – PsbB

Cycle

Temperature (°C)Quantity (ng)

E = 97.9%

R2 = 0.997

∆Rn

Cq

Der

ivat

ive

Rep

(-R

n´)

Yeast – 18S

Cycle

Temperature (°C)Quantity (ng)

E = 90.2%

R2 = 1.000

∆Rn

Cq

Der

ivat

ive

Rep

(-R

n´)

∆Rn

Cq

Melt curveStandard curve

Der

ivat

ive

Rep

(-R

n´)

Mouse kidney – β-actin

Amplification plot

Cycle

Temperature (°C)Quantity (ng)

E = 96.1%

R2 = 0.998

Tobacco leaf – PsbB

Cycle

Temperature (°C)Quantity (ng)

E = 97.9%

R2 = 0.997

∆Rn

Cq

Der

ivat

ive

Rep

(-R

n´)

Yeast – 18S

Cycle

Temperature (°C)Quantity (ng)

E = 90.2%

R2 = 1.000

∆Rn

Cq

Der

ivat

ive

Rep

(-R

n´)

∆Rn

Cq

Melt curveStandard curve

Der

ivat

ive

Rep

(-R

n´)

Mouse kidney – β-actin

Amplification plot

Cycle

Temperature (°C)Quantity (ng)

E = 96.1%

R2 = 0.998

Luna products provide sensitive, accurate detection & quantitation across a wide variety of genomic DNA sources

qPCR targets were quantitated with 50 ng – 0.5 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.

Probe- versus dye-based detection methodsWhich should I choose for my qPCR?

qPCR is typically measured in one of two ways: either an intercalating dye that fluoresces more strongly upon binding to double-stranded DNA, or a fluorescently-labeled “probe” oligonucleotide that anneals to a specific sequence in the PCR amplicon.

Dye-based detection requires only the addition of PCR primers, making it a cost-effective qPCR option. However, the intercalating dye will detect any dsDNA produced in the reaction. Therefore, off-target and non-template amplification (NTC) can be observed for some primer sets, resulting in inaccurate quantitation. Denaturation (melt) curves performed after the PCR can be used to distinguish between correct and nonspecific products. Additionally, only a single amplicon can be measured in a dye-based qPCR with no ability to perform multiplex reactions.

Probe-based detection requires designing and obtaining a sequence-specific fluorescently-labeled probe oligonucleotide in addition to typical PCR primers. This increases assay costs, but probe-based qPCR experiments benefit from extreme specificity and are unlikely to result in inaccurate quantification due to NTC amplification. Multiplex reactions are possible with probes, as different amplicons can be designed with unique fluorophores according to the optical capabilities of the qPCR instrument.

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Optimize your One-Step RT-qPCR with unique WarmStart technologyThe Luna RT-qPCR kits contain a novel, in silico-designed reverse transcriptase (RT) engineered for improved performance. Both the Luna WarmStart Reverse Transcriptase and Hot Start Taq DNA Polymerase, included in these kits, utilize a temperature-sensitive, reversible aptamer, which inhibits activity below 45°C. This enables room temperature reaction setup and prevents undesired non-specific activity. Furthermore, the WarmStart RT has increased thermostability, improving performance at higher reaction temperatures.

Luna RT-qPCR products offer exceptional sensitivity, reproducibility and performance

RT-qPCR targeting human GAPDH was performed using the Luna Universal One-Step RT-qPCR Kit over an 8-log range of input template concentrations (1 μg – 0.1 pg Jurkat total RNA) with 8 replicates at each concentration. Reaction setup and cycling conditions followed recommended protocols, including a 10-minute RT step at 55°C for the thermostable Luna WarmStart Reverse Transcriptase. NTC = non-template control

1 μg

0.1 μg

10 ng

1 ng

0.1 ng

10 pg

1 pg

0.1 pg

Input

NTC

Cycle

∆R

n

Melt curve

Temperature (°C)

Standard curve

De

riva

tive

R

ep

ort

er

(-R

n´)

Quantity (pg)

Cq

Efficiency = 100.4%

R2 = 0.998

10

1

0.12 383634323028262422201810864 40161412

Amplification plot

32.5

25.022.520.017.5

30.027.5

15.012.510.0

0.01 0.1 1 10 102 103 104 105 106 107 65

1.4

1.2

1.0

0.8

0.6

0.4

0.2

0.070 75 80 85 90 95

n = 8

Luna RT-qPCR products offer robust performance in multiplex applications

Multiplex RT-qPCR targeting human GAPDH, ribosomal protein L32g and PI3-Kinase-Related Kinase SMG1 was performed using the Luna Universal Probe One-Step RT-qPCR Kit over a 7-log range of input template concentrations (1 μg – 1 pg Jurkat total RNA) with 4 replicates at each concentration. Amplification plots are shown both overlayed (left) and for each multiplex target (right). To account for copy number differences, 0.4 μM primer was used for lower-copy target (SMG1) and 0.2 μM primer for higher-copy targets (L32g and GAPDH). Luna maintains superior efficiency, reproducibility, sensitivity and performance in multiplex RT-qPCR. NTC = non-template control

∆R

n

1 μg

0.1 μg

10 ng

1 ng

0.1 ng

10 pg

1 pg

Input

Log Quantity (pg)

Cq

Amplification plot

Standard curve

GAPDH (FAM)

L32g (HEX)

SMG1 (Cy5)

Cycle

Cycle

GAPDH (FAM)Efficiency = 99.2%R2 = 1.000

L32g (HEX)Efficiency = 101.5%R2 = 1.000

SMG1 (Cy5)Efficiency = 98.1%R2 = 1.000

NTC

1

0.1

0.01

1

0.1

2 6 10 14 18 22 26 30 34 38 42

Cycle

0.1 1 5 10 30 100 200 1000 10000 100000 1000000 10000000

25

30

35

40

20

15

10

∆R

n

2 6 10 14 18 22 26 30 34 38 42

1

0.01

0.1

2 6 10 14 18 22 26 30 34 38 42

1

0.01

0.1

2 6 10 14 18 22 26 30 34 38 42

Cycle

∆R

n∆

Rn

Luna WarmStart Reverse Transcriptase prevents spurious amplification resulting from room-temperature pre-incubation

RT-qPCR targeting human ribosomal protein L32 was performed before and after a 24-hour incubation at room temperature, with triplicate reactions for a 5-log range of input human (Jurkat) total RNA and a non-template control. The Luna Universal One-Step RT-qPCR Kit featuring Luna WarmStart Reverse Transcriptase exhibited robust performance and no detectible non-template amplification, either without or with a 24 hour, 25°C pre-incubation, while the ABI 1-Step Kit, featuring a non-WarmStart reverse transcriptase, exhibited significant non-template amplification as seen in the respective melting curves.

Melt curves

Amplification plots

Melt curves

Amplification plots

50 ng 0.5 ng Non-template control (NTC)5 ng 50 pg 5 pg

NEB Luna Universal One-Step RT-qPCR Kit(featuring Luna WarmStart Reverse Transcriptase)

ABI Power SYBR® Green 1-Step Kit(featuring ArrayScript™ UP Reverse Transcriptase)

Temperature (°C)Temperature (°C)∆

Rn

Deriva

tive

Report

er

(-R

n´)

Cycle Cycle

AfterBefore

Cycle

∆R

n

Temperature (°C)Temperature (°C)

Deriva

tive

Report

er

(-R

n´)

Cycle

Before After10

1

0.12 6 10 14 18 22 26 30 34 38

10

1

0.12 6 10 14 18 22 26 30 34 38

10

1

0.1

10

1

0.12 6 10 14 18 22 26 30 34 38

2.4

1.9

1.4

0.9

0.4

2 6 10 14 18 22 26 30 34 38

65 70 75 80 85 90 95

2.4

1.9

1.4

0.9

0.4

65 70 75 80 85 90 95

1.5

1.7

1.3

1.1

0.9

0.5

0.7

0.3

0.1

65 70 75 80 85 90 95

1.5

1.7

1.3

1.1

0.9

0.5

0.7

0.3

0.1

65 70 75 80 85 90 95

Melt curves

Amplification plots

Melt curves

Amplification plots

50 ng 0.5 ng Non-template control (NTC)5 ng 50 pg 5 pg

NEB Luna Universal One-Step RT-qPCR Kit(featuring Luna WarmStart Reverse Transcriptase)

ABI Power SYBR® Green 1-Step Kit(featuring ArrayScript™ UP Reverse Transcriptase)

Temperature (°C)Temperature (°C)

∆R

nD

eriva

tive

Re

po

rte

r (-

Rn

´)

Cycle Cycle

AfterBefore

Cycle

∆R

n

Temperature (°C)Temperature (°C)

De

riva

tive

Re

po

rte

r (-

Rn

´)

Cycle

Before After10

1

0.12 6 10 14 18 22 26 30 34 38

10

1

0.12 6 10 14 18 22 26 30 34 38

10

1

0.1

10

1

0.12 6 10 14 18 22 26 30 34 38

2.4

1.9

1.4

0.9

0.4

2 6 10 14 18 22 26 30 34 38

65 70 75 80 85 90 95

2.4

1.9

1.4

0.9

0.4

65 70 75 80 85 90 95

1.5

1.7

1.3

1.1

0.9

0.5

0.7

0.3

0.1

65 70 75 80 85 90 95

1.5

1.7

1.3

1.1

0.9

0.5

0.7

0.3

0.1

65 70 75 80 85 90 95

Melting curves featuring Luna Universal One-Step RT-qPCR Kit before (left) and after (right) 24h pre-incubation at room temperature

Melting curves featuring ArrayScript™ UP Reverse Transcriptase (ABI) before (left) and after (right) 24h pre-incubation at room temperature

Page 7: Luna Universal qPCR & RT-qPCR - New England Biolabs GmbH€¦ · Luna ® Universal qPCR & RT-qPCR LIGHTING THE WAY ™ beINSPIRED driveDISCOVERY stayGENUINE. 2 Fluorescence-based

7

Two-step RT-qPCR uncouples cDNA synthesis and subsequent qPCR, allowing greater freedom in selecting reverse transcriptases (RTs) and qPCR reagents separately. This flexibility can be useful for controlling sequence representation, qPCR efficiency, and optimization of reaction conditions when working with difficult RT-qPCR reactions or low RNA inputs.

The LunaScript RT SuperMix Kit is optimized for first strand cDNA synthesis in the context of a two-step RT-qPCR workflow. It employs an in silico designed Reverse Transcriptase in a convenient SuperMix containing random hexamer and oligo-dT primers, dNTPs and Murine RNase Inhibitor. The kit delivers best-in-class performance and requires the shortest reaction time (< 15 min) and tolerates elevated temperatures (55°C - 65°C), reducing complications from RNA secondary structure.

The cDNA products generated by LunaScript have been extensively evaluated in qPCR using the Luna® qPCR Master Mixes. In combination, these products provide a two-step RT-qPCR workflow with excellent sensitivity and accurate, linear quantitations.

Illuminate your Two-Step RT-qPCR: LunaScript™ RT SuperMix Kit

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

LunaScript™ LunaScriptiScript™SuperScript™ IV

VILO

Dye-based detection Probe-baseddetection

∆Cq

70

80

90

100

110

120

130

Effi

cien

cy (

%)

0 10 20 0 10 20 0 10 20 0 10 20

95%95% 73% 60%Targets passingperformance

criteria

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 the Luna Universal qPCR Master Mix or Luna Universal Probe qPCR Master Mix. 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).

0 10

n = 3

20 30 40102

Cycles

RF

U

0 10 20 30 40

103

Cycles

RF

U

β-actin, human total RNA ERCC00130, ERCC Spike-In Mix 1 RNA

Efficiency: 96.3%

Efficiency: 99.4%

n = 31 µg

100 ng

10 ng

1 ng

100 pg

10 pg

1 pg

NTC

Input

5x109

Input copies

5x108

5x107

5x106

5x105

5x104

5x103

5x102

5x101

NTC

A. B.

0 5 10 15 20 25 30 35 40 45 50 50 60 65 70 75Time (min)

LunaScript™

SuperScript® IV VILO™

Maxima H Minus

iScript™

BioScript™

GrandScript

qScript®

qScript XLT

SuperScript III VILO

95

85

55

50

46

42

25

Temp. (°C)

The LunaScript RT SuperMix Kit offers the shortest available first-strand cDNA synthesis protocol at elevated temperatures.

RNA was converted to cDNA using the 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 the 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) mix 1 RNA containing 5x109 to 50 copies of ERCC00130 (~10 ng – 10 fg) was converted to cDNA and then quantitated by qPCR.

The LunaScript RT SuperMix Kit offers exceptional sensitivity, linearity, and reproducibility in two-step RT-qPCR workflows

The LunaScript RT SuperMix Kit demonstrates superior linear detection of RNA targets

Request a sample at

LUNAqPCR.de (in Germany & Austria)

or LUNAqPCR.fr (in France)

Page 8: Luna Universal qPCR & RT-qPCR - New England Biolabs GmbH€¦ · Luna ® Universal qPCR & RT-qPCR LIGHTING THE WAY ™ beINSPIRED driveDISCOVERY stayGENUINE. 2 Fluorescence-based

One or more of these products are covered by patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc. For more information, please email us at [email protected]. The use of these products may require you to obtain additional third party intellectual property rights for certain applications.

Your purchase, acceptance, and/or payment of and for NEB’s products is pursuant to NEB’s Terms of Sale at www.neb.com/support/terms-of-sale. NEB does not agree to and is not bound by any other terms or conditions, unless those terms and conditions have been expressly agreed to in writing by a duly authorized officer of NEB.

BIORAD® is a registered trademark of BioRad Laboratories. QIAGEN® and QUANTITECT® are registered trademarks of The Qiagen Group. PROMEGA® and GOTAQ® are registered trademarks of Promega Corporation. SSOADVANCED™ is a trademark of BioRad Laboratories. SYBR® is a registered trademark of Molecular Probes, Inc. FASTSTART™ is a trademark of Roche. POWERUP™ and ARRAYSCRIPT™ are trademarks of Thermo Fisher Scientific.

© Copyright 2018, New England Biolabs, Inc.; all rights reserved

Available products include:

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

NEB #M3003S/L/X/E 200/500/1000/2500 rxns

Luna Universal Probe qPCR Master MixRapid, sensitive, and precise probe-based qPCR detection and quantitation of target DNA and cDNA sequences

NEB #M3004S/L/X/E 200/500/1000/2500 rxns

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

NEB #E3005S/L/X/E 200/500/1000/2500 rxns

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

NEB #E3006S/L/X/E 200/500/1000/2500 rxns

Find the right Luna product for your application

NEW LunaScript RT SuperMix KitSingle tube with blue tracking dye includes everything you need for first strand cDNA synthesis in two-step RT-qPCR workflows

NEB #E3010S/L 25/100 rxns

ISO 13485Registered

Medical Devices

ISO 14001Registered

EnvironmentalManagement

ISO 9001Registered

QualityManagement

Selectyour target1

Select yourdetection method2

Genomic DNA or cDNA

Luna Universal ProbeqPCR Master Mix

(NEB #M3004)

Dye-based

Luna UniversalqPCR Master Mix

(NEB #M3003)

RNA

One-Step:Luna Universal

One-Step RT-qPCR Kit(NEB #E3005)

Two-Step:LunaScript Supermix Kit

(NEB #E3010)& Luna Universal qPCR Master Mix

(NEB #M3003)

One-Step:Luna Universal Probe

One-Step RT-qPCR Kit(NEB #E3006)

Two-Step:LunaScript Supermix Kit

(NEB #E3010)& Luna Universal Probe

qPCR Master Mix(NEB #M3004)

Probe-based

CanadaNew England Biolabs, Ltd.Toll Free: [email protected]

China, People’s Republic New England Biolabs (Beijing), Ltd.Telephone: 010-82378265/[email protected]

JapanNew England Biolabs Japan, Inc.Telephone: +81 (0)3 5669 [email protected]

SingaporeNew England Biolabs, PTE. Ltd.Telephone: +65 [email protected]

United KingdomNew England Biolabs (UK), Ltd.Call Free: 0800 [email protected]

Germany & AustriaNew England Biolabs GmbHBrüningstr. 50, Geb B85265926 Frankfurt/Main, GermanyTel: +49/(0)69/305-23140Fax: +49/(0)69/305-23149 Free Call: 0800/246 5227 (Germany)Free Call: 00800/246 52277 (Austria)[email protected]

www.neb-online.deFranceNew England Biolabs France SASGenopole Campus 1, Bâtiment 65 rue Henri Desbruères91030 Evry cedex, FranceTel.: 0800 100 632 (Customer Service)Tel.: 0800 100 633 (Technical Service)FAX.: 0800 100 [email protected]

www.neb-online.frUSANew England Biolabs, Inc.Telephone: (978) 927-5054Toll Free (USA Orders): 1-800-632-5227Toll Free (USA Tech): 1-800-632-7799Fax: (978) [email protected] www.neb.com

www.neb.com

LUNA_TRI – Version 3.0 – 01/19 x/fNEB126