WarmStart® RTx Reverse Transcriptase (Glycerol-free) |NEB酶试剂 New England Biolabs

发表于由

上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。

产品信息

WarmStart RTx Reverse Transcriptase (Glycerol-free) is formulated without glycerol to support lyophilization, incorporation into microfluidic devices, and enable quick adoption into automation workflows.

WarmStart RTx Reverse Transcriptase is a unique in silico-designed RNA-directed DNA polymerase coupled with a reversibly-bound aptamer that inhibits RTx activity below 40°C. This enzyme can synthesize a complementary DNA strand initiating from a primer using RNA (cDNA synthesis) or single-stranded DNA as a template. RTx is a robust enzyme for RNA detection in amplification reactions and is particularly well suited for use in LAMP (Loop-mediated Isothermal Amplification). The WarmStart property enables high throughput applications, room temperature setup, and increases the consistency and specificity of amplification reactions. RTx contains intact RNase H activity.

WarmStart RTx Reverse Transcriptase (Glycerol-free) offers the same robust detection of human RNA targets in RT-LAMP assays as the glycerol-containing enzyme.

WarmStart® RTx Reverse Transcriptase (Glycerol-free) |

RT-LAMP (RNA targets) experiments were performed with NEB #M0380: WarmStart RTx Reverse Transcriptase and NEB #M0538: Bst 2.0 WarmStart DNA Polymerase, or NEB #M0439: WarmStart RTx Reverse Transcriptase (Glycerol-free) and NEB #M0402: Bst 2.0 WarmStart DNA Polymerase (Glycerol-free). Reactions containing 1X LAMP primers and 0.5X LAMP Fluorescent dye were set up in quadruplicate over three logs of total Jurkat RNA (10 ng to 0.1 ng) in 96-well, 25 µl reactions. Control reactions without template (NTC) were also evaluated. Reactions were incubated at 65°C for 40 minutes and fluorescence was monitored every 15 seconds in the SYBR/FAM channel of a real-time thermocycler (Bio-Rad® CFX96). Each dot represents the time at which the fluorescence signal for a single reaction crosses the instrument-defined threshold. All four replicates were detected at each template input unless otherwise indicated (note that dots frequently overlap given similar detection time for the replicates). Overall, similar performance was observed for both glycerol-containing (NEB #M0538 and NEB #M0380) and glycerol-free (NEB #M0402: and NEB #M0439) enzymes at each template input. No amplification was observed in any of the no template control reactions.

产品类别:
Isothermal Amplification & Strand Displacement Products,
cDNA Synthesis & Reverse Transcriptases Products,
PCR, qPCR & Amplification Technologies Products

应用:
Loop-Mediated Isothermal Amplification,
DNA Amplification, PCR & qPCR

  • 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
    • M0439L     -80    
        WarmStart® RTx Reverse Transcriptase (Glycerol-free) M0439LVIAL -80 1 x 0.027 ml 75,000 units/ml
        10X Isothermal Amplification Buffer (Lyo-compatible) B1714SVIAL -20 1 x 1.25 ml 10 X

操作说明、说明书 & 用法

  • 操作说明

    1. Typical RT-LAMP Protocol (NEB #M0439)
    2. Typical cDNA Synthesis Protocol with WarmStart® RTx Reverse Transcriptase (Glycerol-free) (NEB #M0439)

工具 & 资源

  • Web 工具

    • NEB LAMP Primer Design Tool

FAQs & 问题解决指南

  • FAQs

    1. What is LAMP and RT-LAMP?
    2. What advantages does WarmStart® RTx provide?
    3. How sensitive is RNA amplification with WarmStart® RTx?
    4. Can WarmStart® RTx be inactivated?
    5. Is WarmStart® RTx active in other buffers?
    6. What is the maximum length of cDNA product produced by WarmStart® RTx?
    7. Does WarmStart® RTx have RNase H activity?
    8. Can DNA be used as a template for WarmStart® RTx?
    9. What are Hot Start and WarmStart® polymerases and when would I use them?
    10. What is the difference between buffers B1714: 10X Isothermal Amplification Buffer (Lyo-compatible) and B0537: 10X Isothermal Amplification Buffer?
    11. What is the Mg2+ concentration in the 10X Isothermal Amplification Buffer (Lyo-compatible)?
    12. Does B1714: 10X Isothermal Amplification Buffer (Lyo-compatible) include any excipients?
    13. Can WarmStart® RTx Reverse Transcriptase (Glycerol-free) at 75,000 U/mL be diluted prior to use?
    14. How many Freeze/Thaw cycles can WarmStart® RTx Reverse Transcriptase (Glycerol-free) tolerate?

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]

    • M0439L_v1

  • 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#]

    • M0439L_v1_10225699