LunaScript® RT SuperMix Kit | NEB酶试剂 New England Biolabs

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LunaScript® RT SuperMix Kit

  • 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

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

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LunaScript® RT SuperMix Kit | NEB酶试剂 New England Biolabs

Avoiding RNase Contamination

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库存
货号 浓度 规格 目录价 北京 上海 广州 成都 苏州
E3010S 不适用 25 reactions ¥1,379.00
E3010L 不适用 100 reactions ¥4,259.00
如何订购
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LunaScript® RT SuperMix Kit | NEB酶试剂 New England Biolabs Want to try this product? Request a sample LunaScript® RT SuperMix Kit | NEB酶试剂 New England Biolabs Need a custom/large volume order? Contact Us

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产品信息

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

LunaScript® RT SuperMix Kit | NEB酶试剂 New England Biolabs

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

LunaScript® RT SuperMix Kit | NEB酶试剂 New England Biolabs

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

LunaScript® RT SuperMix Kit | NEB酶试剂 New England Biolabs
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 5×109 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

LunaScript® RT SuperMix Kit | NEB酶试剂 New England Biolabs
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

LunaScript® RT SuperMix Kit | NEB酶试剂 New England Biolabs
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 

LunaScript® RT SuperMix Kit | NEB酶试剂 New England Biolabs
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 Kit | NEB酶试剂 New England Biolabs
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.

产品类别:
Luna® qPCR & RT-qPCR Products

应用:
qPCR & RT-qPCR,
Reverse Transcription (cDNA Synthesis)

  • 试剂盒组成

    试剂盒组成

    本产品提供以下试剂或组分:

    NEB # 名称 组分货号 储存温度 数量 浓度
    • 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 不适用
    • 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 不适用

  • 特性和用法

  • 相关产品

    相关产品

    • Luna® Universal qPCR Master Mix
    • Luna® Universal Probe qPCR Master Mix
    • Antarctic Thermolabile UDG
    • Monarch® Total RNA Miniprep Kit

操作说明、说明书 & 用法

  • 操作说明
    1. Protocol for LunaScript® RT SuperMix Kit (E3010)
    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)

  • 说明书
    产品说明书包含产品使用的详细信息、产品配方和质控分析。

    • manualE3010

  • 应用实例
    • Facilitating Detection of SARS-CoV-2 Directly from Patient Samples: Precursor Studies with RT-qPCR and Colorimetric RT-LAMP Reagents

FAQs & 问题解决指南

  • FAQs
    1. Can I use RNA samples purified from different storage conditions such as RNALater in my LunaScript® cDNA synthesis reactions?
    2. Can I add gene-specific primers in my LunaScript® cDNA synthesis reaction?
    3. Can I set up my cDNA synthesis reaction at room temperature when using the LunaScript® RT SuperMix?
    4. Can I use a shorter incubation time for my cDNA synthesis reaction when using the LunaScript® RT SuperMix?
    5. Can I use LunaScript® cDNA products directly in qPCR? How much can I use for detection in a qPCR reaction?
    6. How much RNA template should I use in my LunaScript® cDNA synthesis reactions?
    7. How should I store LunaScript® cDNA products?
    8. How should I choose between a one-step RT-qPCR or two-step RT-qPCR approach?
    9. Should I include a No-Reverse Transcriptase (RT) control reaction when setting up my cDNA synthesis reaction?
    10. What qPCR reagents do you recommend for detection of cDNA products?
    11. What temperature should I use for my LunaScript® cDNA synthesis reaction?
    12. Why do I have low cDNA yields?
    13. Will the blue dye in the LunaScript® RT SuperMix interfere with detection?
    14. How do I choose between the first strand cDNA synthesis kits from NEB?
    15. When using LunaScript cDNA products in downstream PCR (i.e., two-step RT-PCR), what volumes and PCR products can be used?
    16. Can I use LunaScript RT SuperMix in RNA-seq workflows?
    17. How do I choose between LunaScript® RT SuperMix Kit and ProtoScript first strand cDNA synthesis kits?
    18. Can I use a shorter incubation time for my cDNA synthesis reaction when using the LunaScript® RT Master Mix (Primer-free)? Does longer incubation increase the yield of cDNA?

引用 & 技术文献

  • 引用文献

    产品引用文献查找工具

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质控、安全 & 法规

  • 质控分析
    每一新批次的 NEB 产品都要经过质控,以满足指定的质量标准,对特定产品的质量标准和每一批次的检测数据可以通过产品说明表格、COA、产品信息卡或者产品手册进行查阅或下载。关于 NEB 产品质控的详细信息,可从此处查阅。

  • 产品说明与变更通知

    产品说明与变更通知

    产品说明表包含产品的储存温度、有效期和规格,这些文件的命名规则如下:[货号]_[规格]_[版本]

    • E3010S_L_v1
    • E3010S_L_G_v2

  • CoA
    CoA 文件包含单个批次产品的储存温度、有效期和质控,这些文件的命名规则如下: [货号]_[规格]_[版本]_[Lot#]

    • E3010G_v1_10017973
    • E3010L_v1_10019215
    • E3010L_v1_10026112
    • E3010L_v1_10027278
    • E3010L_v1_10031040
    • E3010L_v1_10031843
    • E3010L_v1_10032867
    • E3010L_v1_10036593
    • E3010L_v1_10036597
    • E3010G_v1_10036198
    • E3010L_v1_10041316
    • E3010G_v1_10041336
    • E3010L_v1_10045122
    • E3010S_v1_10040163
    • E3010G_v1_10045186
    • E3010L_v1_10046220
    • E3010L_v1_10047868
    • E3010S_v1_10048557
    • E3010L_v1_10048855
    • E3010S_v1_10051008
    • E3010L_v1_10051006
    • E3010L_v1_10052115
    • E3010S_v1_10054271
    • E3010L_v1_10056224
    • E3010G_v1_10051007
    • E3010G_v1_10059549
    • E3010L_v1_10057893
    • E3010L_v1_10060099
    • E3010S_v1_10059551
    • E3010L_v1_10063504
    • E3010S_v1_10068558
    • E3010L_v1_10068557
    • E3010L_v1_10070738
    • E3010S_v1_10071617
    • E3010L_v1_10072557
    • E3010L_v1_10073018
    • E3010S_v1_10072562
    • E3010L_v1_10074949
    • E3010L_v1_10080307
    • E3010L_v1_10083617
    • E3010L_v1_10086294
    • E3010L_v1_10088901
    • E3010L_v1_10090039
    • E3010L_v1_10092901
    • E3010L_v1_10094979
    • E3010L_v1_10096285
    • E3010S_v1_10096296
    • E3010L_v1_10097139
    • E3010L_v1_10097786
    • E3010L_v1_10098892
    • E3010L_v1_10100557
    • E3010L_v1_10101198
    • E3010S_v1_10101224
    • E3010L_v1_10102449
    • E3010L_v1_10103206
    • E3010L_v1_10104351
    • E3010L_v1_10104814
    • E3010L_v1_10105006
    • E3010L_v1_10105340
    • E3010S_v1_10105341
    • E3010G_v1_10105551
    • E3010L_v1_10106228
    • E3010L_v1_10109591
    • E3010L_v1_10112141
    • E3010G_v1_10110787
    • E3010L_v1_10113203
    • E3010S_v1_10110051
    • E3010L_v1_10114442
    • E3010L_v1_10115719
    • E3010L_v1_10116737
    • E3010L_v1_10117524
    • E3010L_v1_10118873
    • E3010S_v1_10116768
    • E3010G_v1_10119918
    • E3010L_v1_10119618
    • E3010L_v1_10120533
    • E3010L_v1_10121953
    • E3010S_v1_10121265
    • E3010G_v1_10121775
    • E3010L_v1_10122706
    • E3010L_v1_10124311
    • E3010L_v1_10125199
    • E3010L_v1_10126822
    • E3010G_v1_10129004
    • E3010S_v1_10125529
    • E3010S_v1_10128609
    • E3010L_v1_10129544
    • E3010L_v1_10132451
    • E3010L_v1_10135154
    • E3010L_v1_10135691
    • E3010L_v1_10137557
    • E3010L_v1_10138183
    • E3010S_v1_10138428
    • E3010L_v1_10138628
    • E3010L_v1_10140614
    • E3010L_v1_10141536
    • E3010G_v1_10142486
    • E3010L_v1_10143075
    • E3010S_v1_10143235
    • E3010L_v1_10146271
    • E3010G_v1_10149445
    • E3010S_v1_10151712
    • E3010L_v1_10150181
    • E3010G_v2_10152155
    • E3010G_v2_10157115
    • E3010G_v2_10161323
    • E3010L_v2_10155444
    • E3010S_v2_10160093

  • SDS
    以下 SDS 文件可以帮助您安全地使用该产品

    • LunaScript® RT SuperMix
    • No-RT Control Mix
    • Nuclease-free Water