LyoPrime Luna™ Probe One-Step RT-qPCR Mix with UDG |NEB酶试剂 New England Biolabs

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LyoPrime Luna™ Probe One-Step RT-qPCR Mix with UDG

Supplied as a lyophilized cake, the LyoPrime Luna™ Probe One-Step RT-qPCR Mix with UDG enables sensitive detection of target RNA sequences in a room temperature-stable format. This product contains the same versatile features and strong performance as the liquid version: Luna® Probe One-Step RT-qPCR 4X Mix with UDG (NEB #M3019).

  • Simply add nuclease-free water for rapid rehydration (view protocol)
  • Store at room temperature for up to 2 years prior to rehydration
  • Eliminate cold chain shipping requirements
  • Multiplex up to 5 targets to increase throughput
  • Increase reaction specificity and robustness with our unique pairing of Luna WarmStart® RT and Hot Start Taq
  • Prevent carryover contamination with Thermolabile UDG and dUTP included in optimized mix
  • Maintain RNA integrity with Murine RNase Inhibitor included in optimized mix
  • Eliminate pipetting errors with non-fluorescent blue tracking dye
  • Use with a variety of instrument platforms, including those that require a high or low ROX reference signal
  • Developed jointly by New England Biolabs and Fluorogenics™ Limited, which is now a subsidiary of New England Biolabs, Inc.

LyoPrime Luna™ Probe One-Step RT-qPCR Mix with UDG |

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货号 浓度 规格 目录价 北京 上海 广州 成都 苏州
L4001S 不适用 120 reactions ¥2,899.00 有 无 无 无 无
如何订购
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LyoPrime Luna™ Probe One-Step RT-qPCR Mix with UDG | Need a custom/large volume order? Contact us

Bulk packaging may also be available and requested for large recurring orders.
Learn More

产品信息

The LyoPrime Luna Probe One-Step RT-qPCR Mix with UDG is offered in a lyophilized format, allowing for ambient/room temperature shipping and storage prior to use. By simply adding water, the mix can be rehydrated and is ready for use. It is designed for real-time detection of target RNA sequences using hydrolysis probes. Performance in multiplexing applications has been optimized, with sensitive, linear detection of up to 5 targets across a range of inputs. The stable cake can be resuspended to make a 2X or 4X mix to accommodate a variety of sample input volumes. 

Figure 1: LyoPrime Luna offers a robust, versatile RT-qPCR option in a shelf-stable lyophilized format

LyoPrime Luna™ Probe One-Step RT-qPCR Mix with UDG |

The LyoPrime Luna™ RT-qPCR Mix (NEB #L4001) offers the same versatile features as the Luna RT-qPCR 4X Mix (NEB #M3019) in a lyophilized format that can be shipped and stored at room temperature.

The mix contains the necessary components for one-step RT-qPCR, including Luna WarmStart RT, Hot Start Taq DNA Polymerase, dNTPs, and Murine RNase Inhibitor in an optimized buffer. Combining Hot Start Taq DNA Polymerase with a WarmStart-activated reverse transcriptase allows for dual control of enzyme activity via reversible, aptamer-based inhibition. This temperature-dependent activation helps prevent undesirable non-specific priming and extension prior to thermocycling, enabling room temperature reaction set-up after rehydration. The engineered Luna WarmStart RT also possesses higher thermostability than many other RTs, allowing an optimal reaction temperature of 55°C. Additionally, the mix is formulated with a unique passive reference dye that is compatible across a variety of instrument platforms, including those that require a high or low ROX reference signal.

The sensitivity of RT-qPCR makes it important to minimize DNA contamination wherever possible. The inclusion of dUTP and thermolabile UDG prevents carryover contamination, where a product of a previous amplification can serve as the substrate of a subsequent reaction. Unlike E.coli UDG, the thermolabile UDG is completely inactivated at temperatures above 50°C, thereby having no effect on qPCR performance.

A non-fluorescent visible blue tracking dye is included to avoid pipetting errors. The tracking dye does not overlap spectrally with fluorophores commonly used in qPCR and does not interfere with real-time detection.

To learn about the lyophilization process, or to inquire about customizing this product to meet your assay needs, please visit www.neb.com/lyoprime.

Figure 2: Lyophilized and liquid Luna RT-qPCR mixes demonstrate equivalent strong performance

LyoPrime Luna™ Probe One-Step RT-qPCR Mix with UDG |

A. RT-qPCR targeting human β-actin was performed using either the LyoPrime™ Luna Probe One-Step RT-qPCR Mix with UDG (NEB #L4001) or Luna® Probe One-Step 4X Mix with UDG (NEB #M3019) over an 8-log range of input template concentrations (1 μg – 0.1 pg Jurkat total RNA) with 4 replicates at each concentration, run on an ABI® QuantStudio™ 6 Flex real-time instrument. Reactions (20 μl) included primers at 400 nM each and and probe at 200 nM, and followed the product recommended cycling conditions. B. Standard Curve results were substantially equivalent for the lyophilized (gold) and liquid-format (blue) mixes, with strong linearity and reproducibility observed, even at the lowest concentrations tested.
Figure 3: LyoPrime Luna enables robust multiplexing

LyoPrime Luna™ Probe One-Step RT-qPCR Mix with UDG |

Multiplex RT-qPCR was performed using the LyoPrime Luna™ Probe One-Step RT-qPCR Mix with UDG over a 6-log range of Jurkat total RNA (1 μg to 10 pg) on a Bio-Rad® CFX96 real-time instrument. Amplification standard curves and efficiencies for each of the 5 human targets are shown. Reactions (20 μl) included primers and probes at 200 nM each, and followed the product recommended cycling conditions. All five targets were detected linearly in the multiplex reactions with strong efficiency and R2 values.
Figure 4: LyoPrime Luna matches the performance of the liquid Luna reagent and outperforms other commercial lyophilized mixes

LyoPrime Luna™ Probe One-Step RT-qPCR Mix with UDG |

Commercially-available lyophilized RT-qPCR reagents were tested on 8 human RNA targets varying in abundance, length, and %GC. Data was collected by 2 users and according to manufacturers’ recommendations. Results were evaluated for efficiency, low input detection and lack of non-template amplification (where ΔCq = average Cq of non-template control – average Cq of lowest input). In addition, consistency, reproducibility and overall curve quality were assessed (Quality Score). The LyoPrime Luna Probe RT-qPCR Mix with UDG (NEB #L4001) outperformed other lyophilized products, with strong results comparable to those of the liquid format Luna Probe RT-qPCR Mix with UDG (NEB #M3019).

Learn more about our comprehensive qPCR/RT-qPCR testing and “dots in boxes” data visualization here.

Figure 5: Lyophilized and liquid Luna reagents demonstrate sensitive detection of SARS-CoV-2 viral RNA

LyoPrime Luna™ Probe One-Step RT-qPCR Mix with UDG |

Multiplex RT-qPCR was performed using primers and probes from the Luna SARS-CoV-2 RT-qPCR Multiplex Assay Kit (NEB #E3019) targeting 2019-nCoV_N1 (N1, HEX™), 2019-nCoV_N2 (N2, FAM™) and human RNase P (RP, Cy5), run on a Bio-Rad® CFX384 real-time instrument. Reactions (5 μl) included primers at 400 nM each and and probes at 200 nM each, and followed the product recommended cycling conditions. Performance was evaluated using Synthetic Twist SARS-CoV-2 RNA Control 2 diluted in 5 ng/μl human Jurkat total RNA. The Luna and LyoPrime Luna RT-qPCR Mixes both exhibited linear quantitation (standard curves of 100,000 to 10 copies SARS-CoV-2 RNA, left) and sensitive detection (100% detection of N1 and N2 with 40 replicates at 5 and 10 copies SARS-CoV-2 RNA; no false positive detection with 28 negative control replicates). Similar results were obtained for 20 μl reactions in 96-well format.

Figure 6: LyoPrime Luna reagents enable strong detection and quantitation of various infectious disease targets

LyoPrime Luna™ Probe One-Step RT-qPCR Mix with UDG |

Duplex RT-qPCR was performed using primers and probes targeting the indicated viral RNA gene (FAM) and human RNase P (Cy5) on a Bio-Rad CFX96 instrument. Performance was evaluated for triplicate standard curves of 100,000 to 10 copies ATCC® synthetic viral RNA diluted in 5 ng/μl human (Jurkat) total RNA (Chikungunya Virus: VR-3246SD; Dengue-2 Virus: VR-3229SD; Zika Virus: VR-3252SD). For each target the corresponding ATCC reference primer and probe sequences were used. Reactions (20 μl) included primers at 400 nM each and probes at 200 nM and followed the product recommended cycling conditions. The LyoPrime Luna RT-qPCR Mix exhibited linear quantitation and sensitive detection for each target.
产品类别:
Luna® qPCR & RT-qPCR Products

应用:
qPCR & RT-qPCR,
DNA Amplification, PCR & qPCR

  • 产品组分信息

    产品组分信息

    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
    • L4001S     15 to 25    
        LyoPrime Luna™ Probe One-Step RT-qPCR Mix with UDG L4001SVIAL 15 to 25 2 x 60 reactions 不适用

  • 特性和用法

  • 优势和特性

    Features

    • Can be reconstituted at either 2X or 4X if desired to permit higher sample volumes
    • Same versatile features and strong performance of the Luna Probe One-Step RT-qPCR 4X Mix with UDG (NEB #M3019) in a lyophilized format
    • Includes aptamer technology to control RT and DNA polymerase enzyme activity
     

  • 相关产品

    相关产品

    • Nuclease-free Water
    • Luna® Probe One-Step RT-qPCR 4X Mix with UDG
    • Luna® Probe One-Step RT-qPCR 4X Mix with UDG (No ROX)
    • Monarch® Total RNA Miniprep Kit

操作说明、说明书 & 用法

  • 操作说明
    1. Protocol for LyoPrime Luna One-Step RT-qPCR Mix with UDG (NEB #L4001)

  • 应用实例
    • Key considerations for optimal lyophilized reagent development
    • Optimized conditions for the CDC Influenza SARS-CoV-2 (Flu SC2) Multiplex Assay using Luna One-Step RT-qPCR Reagents

FAQs & 问题解决指南

  • FAQs
    1. How should the LyoPrime Luna mix be stored?
    2. How do I rehydrate the LyoPrime Luna mix? Can it be resuspended at a higher concentration?
    3. Are reaction conditions the same for lyophilized and liquid Luna mixes?
    4. When I dissolve the lyophilized product, the solution at first appears turbid. Is this normal?
    5. Can I set up my Luna® RT-qPCR at room temperature?
    6. Why is the Luna® qPCR Mix blue? Will this dye interfere with detection?
    7. Can I run the Luna® qPCR Mix on my qPCR instrument?
    8. Do I need to add ROX?
    9. What RNA samples can be used in RT-qPCR with the Luna® Mix?
    10. How much RNA template should I use in my RT-qPCR reaction?
    11. Can I run multiplex reactions with the Luna® Probe One-Step RT-qPCR Kits? Do I need to change my reaction conditions?
    12. Can I use a ROX-labeled probe with the Luna® Probe Mix that contain a reference dye?
    13. Other companies recommend a 2-min incubation at 25°C for carryover prevention, is a 30-sec incubation at 25°C sufficient?
    14. Can I use the Luna Probe One-Step RT-qPCR 4X Mix with UDG (NEB #M3019) product for viral detection?
    15. What are the differences between the Luna One-Step RT-qPCR products (NEB #E3006, #E3007 and #M3019)?

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]

    • L4001S_v1

  • 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#]

    • L4001S_v1_10133633

  • SDS
    以下 SDS 文件可以帮助您安全地使用该产品

    • LyoPrime Luna™ Probe One-Step RT-qPCR Mix with UDG