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进口试剂

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RapiDxFire Thermostable Reverse Transcriptase, 250 Rxns

进口试剂

RapiDxFire Thermostable Reverse Transcriptase, 250 Rxns

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产品名称:

RapiDxFire Thermostable Reverse Transcriptase, 250 Rxns

货号
30250-2
规格
250 Rxns
存储温度
没有此类产品
产品描述

RapiDxFire™ Thermostable Reverse Transcriptase

一种真正的热稳定逆转录酶,用于快速合成短cDNA<1 Kb

· 在高温(55-80°C)下极其活跃:通过各种RNA模板提高cDNA合成的特异性

· 敏感: 在两步RT-qPCR分析中检测≤100拷贝的RNA

· 反应时间短(5分钟或更短): 简化的RT-qPCR工作流程和更快的出结果时间

· 在室温下稳定(> 3个月):简化自动化卡座上的设置(无需冷藏),可在冷藏有限的环境中使用

· Lyo兼容: 酶制剂不含甘油和其他成分,不会干扰下游冻干步骤

· 批次间重复性:ISO 13485认证的设施中制造

产品包含RapiDxFire Thermostable Reverse Transcriptase3 Units /μL)和10X RapiDxFire Reaction Buffer。酶包装大小(50250 Rxns)基于25μL反应体积

 

RapiDxFire™热稳定逆转录酶是一种显着优于流行的MMLVAMV逆转录酶的方法。最初在温泉中发现,这种强大的噬菌体酶在升高的温度下表现出越来越高的活性(在9010分钟后活性保持约60%)并且在20-25下储存> 3个月后保持其核心活性。这种酶缺乏RNAse H3'→5'核酸外切酶活性并有效合成短cDNA片段(≤1Kb)。与其他逆转录酶一样,RapiDxFire RT具有DNA聚合酶活性,但缺乏5'→3'核酸外切酶活性。酶的储存缓冲液不含甘油和Triton™X-100可进一步优化用于下游冻干步骤。

RapiDxFire耐高温RT在高温下表现最佳

Figure 1. Reaction temperature profile for RapiDxFire Thermostable Reverse Transcriptase. Reverse transcription reactions were set up on ice using each manufacturer’s recommended buffer system and Poly (rC) /p(dG)12-18 template/primer substrate. Reactions were transferred from ice to the indicated temperatures (37, 50, 55, 60, 65, 70, 75, and 80°C) and incubated for 40 minutes. Following incubation, RNA/cDNA heteroduplex product is quantified as a measure of polymerization activity, utilizing PicoGreen® fluorescence on a Tecan Infinite® M1000 Pro. RapiDxFire Thermostable RT exhibits increasing activity as the reaction temperature is increased, up to 80ºC (highest temperature tested); ~60% activity remains after 10 min at 90⁰C (data not shown).


Superior Detection of Zika virus with RapiDxFire Thermostable RT

RapiDxFire Thermostable RT优异检测寨卡病毒

 

Figure 2. cDNA synthesis time course studies in a 2-step RT-qPCR reaction with different thermostable RTs. A) qPCR curve after a 1 minute reverse transcription reaction. Zika cDNA synthesis was conducted for each enzyme in duplicate using target-specific primers and recommended reaction buffer and incubation temperatures per suppliers’ guidelines (RapiDxFire= 60⁰C, Supplier B= 50⁰C, and Supplier T= 55⁰C). After cDNA synthesis real time PCR was performed using one-tenth volume of each of the cDNA samples. PCR was performed using EconoTaq and its supplied buffer (Lucigen Cat No. 30031-3). Detection of PCR products were done real time using an intercalating dye (Dyomics Cat No. V13-01184) and BioRad CFX C1000 Touch™ with absorption/emission of 481nm/526nm. B) Reverse transcription time course study prior to performing second-step real-time PCR. Encircled data points are derived from data represented in the qPCR curve in A).


3个月室温稳定性


3. RapiDxFire热稳定性室温环境和-20°C下每个时间点单独存储活性对比。 在每个时间点RapiDxFire Thermostable RT通过测量10000拷贝MS2 RNA的第一链cDNA合成来评估。使用MS2的基因特异性引物在60的反应温度下进行cDNA合成5分钟。使用实时PCR和围绕520碱基对扩增子设计的MS2引物测量cDNA

产品名称

规格

货号

RapiDxFire Thermostable Reverse Transcriptase, 3 Units/µL

 

50 Rxns

30250-1

 

 

250 Rxns

30250-2

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