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通用型一步法RT-PCR试剂盒说明书前言本试剂盒适用于对各种动、植物、病毒RNA进行PCR 检测。
用户可根据被分析基因，配合一对引物，或同时采用Taqman 荧光探针，先将提取的RNA反转录(reverse transcription).成cDNA，再经过聚合酶链式反应(Polymerase Chain Reaction)技术对特异性片段进行扩增，进行电泳或实时荧光分析。
一步法RT-PCR可使RT及PCR过程在单管中完成，比分开进行RT及PCR更方便。
由于不需要在RT完成后打开反应管，尽可能地避免了样品间的相互污染。
规格20人份适用仪器适用于各种普通PCR仪器和实时荧光PCR仪器。
试剂盒组成试剂准备根据下表配制反应液：振荡混匀后，按每管 40 μl(可根据需要调整)分装，备用。
PCR扩增将均一化后的RNA提取样本10 μl加入各反应管，混匀、离心后，置普通PCR仪器或实时荧光PCR仪器上进行扩增及实时检测。
结果判断扩增产物可直接进行电泳检测；实时荧光检测可根据相应仪器的配套软件进行结果分析。
保存及有效期-20℃保存，有效期为6个月。
注意事项1. 开始检测前请仔细阅读本说明书全文。
2. 整个检测过程中，反应体系的配制、样本处理及加样、PCR扩增（荧光检测）应分区进行以避免污染。
3. 操作人员应戴口罩，经常更换一次性手套，以避免RNA酶的污染；实验中所用器具均应经过除RNA酶处理。
4. 试剂盒组成中的试剂使用前应充分融化并混匀。
5. 进行实时荧光分析时，应使用透光性能较好的一次性薄壁离心管；.荧光探针应避光保存，加入缓冲液中后，应尽快进行扩增。
6. 注意适当处理检测中遗留的样品、扩增产物及可能被污染的试剂。
生产企业：上海蓝创生物科技发展有限公司。
Code No. 3732 研究用CellAmp™Direct RNA Prep Kitfor RT-PCR (Real Time)说明书目录内容页码●制品说明 1 ●制品内容 1 ●保存 1 ●试剂盒外必备主要试剂 1 ●使用注意 1 ●操作方法 2 ●附录 3 ●实验例9 ● Troubleshooting 10 ●关联产品10●制品说明本制品是从96孔板或其他各种培养板中培养的动物细胞中提取One Step或Two Step Real Time RT-PCR （又称qRT-PCR或RT-qPCR）反应用的RNA模板的试剂盒。
本试剂盒由三种溶液组成，快捷方便，操作简单，可在10分钟内完成从培养细胞中制备品质良好的RNA模板。
使用本试剂盒制备的RNA模板可与One Step TB Green® PrimeScript™RT-PCR Kit (Code No. RR066A/B) 等One Step Real Time RT-PCR试剂组合使用，2小时内便可完成基因表达分析。
本试剂盒与反转录试剂PrimeScript RT reagent Kit（Perfect Real Time）组合使用，可在30分钟内制备出用于Real Time PCR (qPCR) 的cDNA模板。
本试剂盒可以从微量的细胞中提取RNA模板，并用于基因解析和Real Time RT-PCR的高灵敏度检出。
在对无法设计跨内含子引物的基因或低表达量的基因进行分析时，基因组DNA的混入对实验结果影响很大。
本试剂盒可以有效去除基因组DNA，大大提高了实验结果的准确性。
●制品内容（200次量）*CellAmp Washing Buffer 12.5 ml×2CellAmp Processing Buffer 10 mlDNase I for Direct RNA Prep 200 μl*使用96孔板时，200孔培养细胞的反应次数。
一步法RT-PCR 检测试剂盒 (一步法RT-PCR 扩增试剂盒)(目录号HS0612-2)产品包装保存条件-20℃产品简介本试剂盒是专为一步法RT-PCR 实验研制，逆转录和PCR 在同一反应体系中进行，反应过程中无需添加试剂，无需打开管盖，在避免污染的同时提高了检测灵敏度和实验效率。
本试剂盒包括全新高效逆转录酶、快速热启动DNA 聚合酶，同时包含适用于逆转录和PCR 扩增的反应缓冲液和实验中所必需的反应组分。
经过重组表达的SuperRT 逆转录酶RNase H 活性缺失，减少了逆转录反应中RNA 的降解，更容易获得全长的cDNA 。
该逆转酶逆转录效率高，可对少量 RNA 模板进行良好的逆转录反应。
SuperRT 逆转录酶与RNA 亲合性高，能通读GC 含量高，二级结构复杂的RNA 模板，获得高产量的cDNA 。
PCR 反应使用的DNA 聚合酶具有扩增效率高、延伸速度快的优良性能。
独特的缓冲体系使逆转录酶和聚合酶同时发挥最大功效。
使用该试剂盒能够方便快捷的在同一个反应管内完成逆转录和PCR 扩增反应。
使用本试剂盒扩增得到的目的产物3′端附有一个″A ″碱基，可直接用于T/A 克隆。
北京厚生博泰科技有限公司Beijing Hooseen Biotech Co., Ltd.产品特点1. 效率高，可以高效转录GC含量高，二级结构复杂的RNA模板。
2. 耐热性及稳定性强。
3. 操作简单迅速,最大限度的避免污染。
4. 独特的缓冲体系使逆转录酶和聚合酶同时发挥最大功效。
使用方法1）一般PCR实验中退火温度比扩增引物的熔解温度Tm低5℃，退火时间一般为20 ~ 30秒，无法得到理想的扩增效率时，适当降低退火温度；发生非特异性反应时，提高退火温度，由此优化反应条件。
2）延伸时间根据所扩增的片段大小设定，本产品中所包含的DNA Polymerase的扩增效率为1 kb/30s。
3）可根据扩增产物的下游应用设定循环数。
Code No.：DRR041ASYBR® Premix Ex TaqTM （Perfect Real Time） (200 次量)目内 容●制品说明 ●制品内容 ●适用的 Real Time PCR 扩增仪 ●保 存录页 码1 1 1 1 1 1 2 2 2 2 3 3 3●TaKaRa Ex TaqTM HS活性定义 ●纯 度●Real Time PCR 性能检测 ●试剂盒原理 ●试剂盒特长 ●操作注意 ●Real Time PCR 操作顺序 ●操作方法 ◆应用 Thermal Cycler Dice Real Time System 扩增仪的操作方法 ◆应用 ABI PRISM 7000/7700/7900 HT， 7300/7500/7500 Fast Real - Time PCR 的操作方法 ◆应用 Light Cycler Real Time PCR 扩增仪的操作方法 ◆应用 Smart Cycler II System Real Time PCR 扩增仪的操作方法 ◆应用 Mx3000P Real Time PCR 扩增仪的操作方法 ●PCR 反应条件说明 ●实验例 ●进行 RT-PCR 反应时的实验方法 ●引物设计说明 ●引物设计服务说明： 本公司提供用于基因表达定量分析的引物设计及合成服务 ●问 答4 5 7 8 9 9 11 1314 14◆特别提示： 本制品请于 4℃避光保存，避免冻结制品！●制品说明本制品是采用SYBR® Green I嵌合荧光法进行Real Time PCR的专用试剂。
制品中已经将DNA聚合酶、反 应用Buffer、dNTP、SYBR® Green I等试剂预混在一起，是一种 2×浓度的Premix Type试剂，进行实验 时，PCR反应液的配制十分方便简单。
制品中的DNA聚合酶使用了改良后的Hot Start法用 DNA聚合酶TaKaRa Ex TaqTM HS，与TaKaRa精心研制的Real Time PCR用Buffer组合使用，可以有效抑制非特异性的PCR扩增，大大提高PCR的扩增效率，进行高灵敏度的Real Time PCR扩增反应。
Product Components List Takara Bio USA, Inc.1290 Terra Bella Avenue, Mountain View, CA 94043, USA U.S. Technical Support: ********************United States/Canada 800.662.2566 Asia Pacific+1.650.919.7300Europe+33.(0)1.3904.6880Japan+81.(0)77.565.6999Page 1 of 1(050319) Lenti-X™ qRT-PCR Titration KitCatalog No. Amount631235 200 rxnsDescriptionThis Lenti-X qRT-PCR Titration Kit and a real-time PCR instrument can be used to rapidly quantify the virion content of HIV-1-based lentiviral supernatants. The viral genome copy number in a supernatant sample is determined by first purifying the viral RNA, and then comparing the RT-PCR threshold cycle (C t) of an appropriate dilution of purified viral RNA to sample curve C t values that are derived from a calibrated, serially diluted control viral RNA template.Package Contents•Lenti-X qRT-PCR Titration Components (Cat. No. 631236; Not sold separately) >>View Components•NucleoSpin RNA Virus Kit (Cat. No. 740956.10)•Quant-X™ One-Step qRT-PCR TB Green® Kit (200 rxns) (Cat. No. 638317; Not sold separately) >>View ComponentsFor storage conditions, please see the Certificate of Analysis supplied with each component.Product DocumentsDocuments for our products are available for download at /manualsThe following documents apply to this product:•Lenti-X qRT-PCR Titration Kit User Manual•Lenti-X qRT-PCR Titration Kit Protocol-at-a-Glance•Viral RNA Isolation User Manual•Quant-X One-Step qRT-PCR TB Green Kit User Manual•Quant-X One-Step qRT-PCR TB Green Kit Protocol-at-a-GlanceNotice to PurchaserOur products are to be used for research purposes only. They may not be used for any other purpose, including, but not limited to, use in drugs, in vitro diagnostic purposes, therapeutics, or in humans. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without prior written approval of Takara Bio USA, Inc.Your use of this product is also subject to compliance with any applicable licensing requirements described on the product’s web page at . It is your responsibility to review, understand and adhere to any restrictions imposed by such statements.© 2009 Takara Bio Inc. All Rights Reserved.All trademarks are the property of Takara Bio Inc. or its affiliate(s) in the U.S. and/or other countries or their respective owners. Certain trademarks may not be registered in all jurisdictions. Additional product, intellectual property, and restricted use information is available at .This document has been reviewed and approved by the Quality Department.。
Code No. RR047A 研究用PrimeScript™RT reagent Kitwith gDNA Eraser(Perfect Real Time)说明书目录内容页码●制品说明1●制品内容 1 ●试剂盒外必备材料 1 ●保存 1 ●特长 2 ●使用注意 2 ●操作方法 2 ● Real Time PCR 4 ●实验例 6 ●附录7 ●关联产品8●制品说明为了准确地进行基因表达量分析，必须满足只有cDNA作为模板检出的先决条件，但Total RNA中常常混有基因组DNA，并可以直接作为PCR反应的模板进行扩增，因此会造成解析结果不准确。
为了避免这种情况发生，通常将检测用引物设计在内含子前后的外显子上，使基因组DNA得不到扩增。
但是，此方法不适合具有单个外显子的基因或两个外显子之间所跨的内含子过小的基因，同时当基因组上有伪基因存在时、或设计引物对基因组有非特异性扩增时、以及基因信息没被完全解析的生物种等也同样不适合于本方法。
在这种情况下，我们常常需要对Total RNA样品进行DNase I处理，以除去残存的基因组DNA。
而DNase I处理通常要进行复杂的纯化操作，同时会造成RNA的降解和损失。
PrimeScript RT reagent Kit with gDNA Eraser是可以除去基因组DNA进行Real Time RT-PCR反应的专用反转录试剂。
Kit中使用了具有较强DNA分解活性的gDNA Eraser，通过42℃，2 min即可除去基因组DNA。
同时由于反转录试剂中含有抑制DNA分解酶活性的组分，经过gDNA Eraser处理后的样品可以直接进行15 min的反转录反应合成cDNA，因此，20 min内即可迅速完成从基因组DNA去除到cDNA合成的全过程。
使用本制品合成的cDNA适用于SYBR® Green分析法和TaqMan®探针分析法，可以根据实验目的，选择与SYBR Premix Ex Taq™ II（Tli RNaseH Plus）（Code No. RR820Q/A/B）、Probe qPCR Mix（Code No. RR391S/A/B）组合使用。
How to Avoid the Generation of Nonspecific Amplification When Using TaqMan Assays-on-Demand Gene Expression ProductsData SheetIntroductionA common concern for real-time PCR users is reactions that produce low amplification efficiency. A reaction with low amplification efficiency will result in low reproducibility ofreplicates, later or delayed Cts for lower DNA input amounts, and lower limits of detection. Pre-designed assays (like AOD) are a convenient method for testing expression levels of known genes. Some AOD targets can give rise to amplification of secondary non-specific PCR products leading to erroneous data analysis and misinterpretation of expression profiles due to decreased sensitivity of detection. The results suggest that having a more robust chemistry and superior hot start technology, as employed in Brilliant III Ultra-Fast, can improve the specificity of amplificationfor these problematic primer and probe sets.In validating Brilliant III Ultra-Fast QPCR and QRT-PCR Master Mixes over many targets and a broad template dilution range, some TaqMan ‘Assays-on Demand’ (AOD) generated primer-dimers with a competitor’s fast QPCR master mix. Such PCR artifacts were not observed with the Brilliant III Ultra-Fast reagents, suggesting the novel hot start technology of Brilliant III prevents generation of non-specific secondary products, providing the user with a greater degree of confidence in their results.2Experimental MethodsTotal RNA from Stratagene Universal Human Reference RNA (cat. # 740000) was reverse transcribed using a mix of oligo(dT) and random primers (AffinityScript QPCR cDNA Synthesis kit; cat. # 600559) to generate cDNA. PCR assays (20µl) contained 1x primer/probe mix (GUS or TBP Assays-on-Demand) and cDNA concentrations ranging from 100-0.01 ng per reaction. Thermal cycling was performed on the ABI StepOnePlus Real-Time PCR system (see Figures 1 & 3 for details of cycling parameters for Brilliant III Ultra-Fast QPCR (cat. # 600880) Master Mix andCompetitor A master mix). The resulting PCR products (5 μL) were evaluated on an Agilent Bioanalyzer 2100 instrument using a DNA 1000 Chip (cat. # 5067-1504).ResultsBrilliant III Ultra-Fast QPCR Master Mix was compared against a commercially available master mix (Competitor A) on two TaqMan AOD gene expression products (GUS and TBP) over a 4-log dilution range (100, 10, 1, 0.1, 0.01 ng/Rxn). Figures 1 & 3 (A &B) show the amplification plots and standard curves of the two fast QPCR master mixes with the GUS and TBP gene expression products respectively. The results indicated a significant difference in amplification efficiencies between the two master mixes on both AOD targets, suggesting further analysis was required to determine the nature of the disparity.Figure 1.Amplification plots and corresponding standard curve of GUS primer/probe set (ABI Assays-On-Demand) using (A) Brilliant III Ultra-Fast Master Mix and (B) using Competitor A master mix on the StepOnePlus PCR system.Reactions (20ul) contained 1x primer/probe and human universal cDNA ranging from 100-0.01 ng/Rxn. Thermal cycling conditions for Brilliant III Ultra-Fast Master Mix were 3min at 95°C initial denaturation followed by 40 cycles of 5sec at 95°C and 10sec at 60°C. Thermal cycling conditions for Competitor A master mix were 20sec at 95°C initial denaturation followed by 40 cycles of 1 sec at 95°C and 20 sec at 60°C. The standard curves span four orders of magnitude resulting in an Rsq of 0.999 and amplification efficiency of 94.3% for the Brilliant III Ultra-Fast Master Mix and an Rsq of 0.996 and amplification efficiency of 79.4% for Competitor A master mix.D R n2.502.252.001.751.501.201.000.750.500.250.002 6 10 14 18 22 26 30 34 38 40CycleAmplification Plot3634323028262422.001 .002 .01 .02 .1 .2 1 2 345 10 20 30 100 200 1000Quantity Target Target 1 GUS Slope -3.466 Y-Inter 28.746 R 0.999 Eff%94.316: : :::C TStandard Curve1.51.41.31.21.11.0.9.8.7.6.5.4.3.2.1.0-.12 6 10 14 18 22 26 30 34 38 40CycleAmplification PlotD R n38363432302826242220.001 .002 .01 .02 .1 .2 1 2 345 10 20 30 100 200 1000Quantity Target Slope Y-Inter R Eff%C TStandard Curve3Figure 2.Trace of GUS qPCR products generated using Competitor A master mix (left) or Brilliant III Ultra-Fast Master Mix (right) run on Agilent 2100 Bioanalyzer. Presence of primer-dimers/non specific amplification is evident when Competitor A master mix is used for amplification.1.81.71.51.31.10.90.70.50.30.1-0.12 6 10 14 18 22 26 30 34 38 40CycleAmplification PlotD R n3735333129272523.001 .002 .01 .02 .1 .2 1 2 3 45 10 2030 100 200 1000Quantity C TStandard CurveTarget Target 2 TDP Slope -3.328 Y-Inter 30.396 R 0.998 Eff%99.735: : :::39383634323028262422QuantityC TStandard Curve.001 .002 .01 .02 .1 .2 1 2 3 45 10 2030100 200 1000Target Target 2 TBP Slope -3.73 Y-Inter 29.979 R 0.993 Eff%85.391: : :: :2.001.002 .01 .02 .1 .21 2 34 5 10 20 30 100 200 10002 6 10 14 18 22 26 30 34 38 40CycleAmplification Plot1.51.31.10.90.70.50.30.1-0.1D R nFigure 3.Amplification plots and corresponding standard curve of TBP primer/probe set (ABI Assays-On-Demand) using (A) Brilliant III Ultra-Fast master mix and (B) using Competitor A master mix on the StepOnePlus PCR system.Reactions (20ul) contained 1x primer/probe and human universal cDNA ranging from 100-0.01 ng/Rxn. Thermal cycling conditions for Brilliant III Ultra-Fast master mix were 3min at 95°C initial denaturation followed by 40 cycles of 5sec at 95°C and 10sec at 60°C. Thermal cycling conditions for Competitor A master mix were 20sec at 95°C initial denaturation followed by 40 cycles of 1sec at 95°C and 20sec at 60°C. The standard curves span four orders of magnitude resulting in an Rsq of 0.998 and amplification efficiency of 99.7% for the Brilliant III Ultra-Fast master mix and an Rsq of 0.993 and amplification efficiency of 85.4% for Competitor A master mix.In order to investigate the cause for the amplification efficiency discrepancy, all amplification products were run on an Agilent Bioanalyzer (Figures 2 & 4). Non-specific PCR products of low molecular weight (primer-dimers) were observed for Competitor A’s fast master mix with both AOD targets. Not surprisingly, the amount of these non-specific products increases as the amountSecondary non-specific product Competitor ABrilliant III Ultra-Fast Master MixSecondary non-specific productCompetitor ABrilliant III Ultra-Fast Master MixFigure 4.Trace of TBP qPCR products generated using Competitor A (left) or Brilliant III Ultra-Fast Master Mix (right) run on Agilent 2100 Bioanalyzer.Presence of primer-dimers/non specific amplification is evident when Competitor A master mix is used for amplification.ConclusionNon-specific amplification and primer-dimer formation can greatlyreduce amplification efficiency of the target gene since theycompete for reaction components during amplification, resultingin later Cts, lower sensitivity, decreased replicate reproducibility,and reduced dynamic range. These performance attributes maydrastically impact the end result. Pre-designed assays, such asAOD, have the potential to generate primer-dimers and throwinto doubt the validity of the analysis. The data presented hereDescription Quantity Reactions*Catalog No. Brilliant III Ultra-Fast QPCR Master Mix for ABI StepOnePlus and BioRad CFX96 2 x 2 ml400600880 Brilliant III Ultra-Fast QPCR Master Mix for ABI StepOnePlus and BioRad CFX (10 pack)20 x 2 ml4000600881 Brilliant III Ultra-Fast QRT-PCR Master Mix for ABI StepOnePlus and BioRad CFX96 2 x 2 ml400600884 Brilliant III Ultra-Fast QRT-PCR Master Mix for ABI StepOnePlus and BioRad CFX96 (10 pack)20 x 2 ml4000600885 Brilliant III Ultra-Fast SYBR Green QPCR Master Mix for ABI StepOnePlus and BioRad CFX96 2 x 2 ml400600882 Brilliant III Ultra-Fast SYBR Green QPCR Master Mix for ABI StepOnePlus and BioRad CFX (10 pack)20 x 2 ml4000600883 Brilliant III Ultra-Fast SYBR Green QRT-PCR Master Mix for ABI StepOnePlus and BioRad CFX96 2 x 2 ml400600886 Brilliant III Ultra-Fast SYBR Green QRT-PCR Master Mix for ABI StepOnePlus and BioRad CFX96 (10 pack)20 x 2 ml4000600887Learn more about Brilliant III Ultra-Fast QPCR Master Mixes and how they can improve your quantification on any Real-Time PCR instrument at /brilliant3.Find an Agilent customer center in your country:/chem/contactsU.S. and Canada1-800-227-9770*****************************Asia Pacific************************Europe************************This item is intended for Research Use Only. Not for use in diagnostic procedures. Information, descriptions and specifications in this publication are subject to change without notice.Agilent Technologies shall not be liable for errors contained herein or for incidental or consequential damages in connection with the furnishing, performance or use of this material.SYBR® is a registered trademark of Molecular Probes.© Agilent Technologies, Inc. 2010Printed in the USA, February 1, 20105990-5401ENillustrates the importance of the QPCR master mix on the final result. The novel Taq mutant and new hot start technology of Brilliant III Ultra-Fast Master Mixes reduces the need for more extensive assay validation and provides more reliable and consistent data across a wider range of different assays.The versatile Brilliant III Ultra-Fast QPCR reagents offer the greatest flexibility in master mixes delivering superior performance across an array of quantitative applications on any Real-Time PCR platform.。
Code No. RR036A 研究用PrimeScript™RT Master Mix(Perfect Real Time)说明书v201605Da目录内容页码●制品说明1●制品内容 1 ●试剂盒外必备材料 1 ●保存 1 ●特长 1 ●使用注意 2 ●操作方法 2 ● Real Time PCR 2 ●实验例 5 ●附录 5 ●关联产品7●制品说明本制品是2 Step Real Time RT-PCR用的理想反转录反应试剂。
5×PrimeScript RT Master Mix中含有定量RT-PCR的反转录反应所需的所有试剂（PrimeScript RTase、RNase Inhibitor、Random 6 mers、Oligo dT Primer、dNTP Mixture、反应Buffer），加入模板RNA和水即可迅速进行反应。
使用具有较强延伸能力的PrimeScript RTase，与制品PrimeScript RT reagent Kit（Perfect Real Time）（Code No. RR037Q/A/B）相同，可以在较短时间内高效合成Real Time PCR用模板cDNA。
本制品合成的cDNA可以用于SYBR® Green qPCR分析法和探针分析法，可以根据实验目的与SYBR Premix Ex Taq™II (Tli RNaseH Plus)（Code No. RR820Q/A/B）、SYBR Fast qPCR Mix（Code No. RR430S/A/B）和Probe qPCR Mix（Code No. RR391S/A/B）等定量试剂配合使用，能够进行高性能的基因表达分析。
注意：Takara Bio使用SYBR Green I作为研究试剂已得到Molecular Probes Inc.的许可。
●制品内容（10 μl反应×200次）1. 5×PrimeScript RT Master Mix（for Real Time）*1 400 μl2. RNase Free dH2O 1 ml×2支3. EASY Dilution（for Real Time PCR）*2 1 ml*1：含有PrimeScript RTase、RNase Inhibitor、Random 6 mers、Oligo dT Primer、dNTP Mixture、反应Buffer（含有Mg2+）。
Catalog No. RT0411-01产品简介2×SYBR Green Mix（With ROX）是使用染料法（SYBR GreenⅠ）进行实时荧光定量PCR扩增反应的预混体系，包括Hotstart Taq DNA Polymerase，Buffer，dNTPs，SYBR Green I荧光染料、Mg2+和ROX校正染料。
本品含有经化学修饰的全新高效的热启动酶Hotstart Taq DNA Polymerase，在常温下没有聚合酶活性，有效避免在常温条件下由引物和模板非特异性结合或引物二聚体而产生的非特异性扩增，酶的激活须在95℃下孵育10 分钟。
主要用于基因组DNA靶序列和RNA反转录后cDNA靶序列的检测。
本品所含的荧光染料SYBR Green I可以与所有的双链DNA结合，使该产品可用于不同靶序列的检测而不需合成特异性标记探针。
所含的ROX染料可校正定量PCR仪孔与孔之间产生的荧光信号误差, 适用于以ROX作为校正染料的所有荧光定量PCR仪。
产品包装保存条件-20℃避光保存，尽量避免尽量避免反复冻融。
注意事项1 使用前请上下颠倒轻轻混匀，尽量避免起泡，并经短暂离心后使用。
2 本产品中含有SYBR Green I 荧光染料和ROX 染料，保存本产品或配制PCR 反应液时应避免强光照射。
3 避免反复冻融本品，反复冻融可能使产品性能下降。
如果在短期内需要频繁使用，可在2-8℃保存。
使用方法以下举例为常规PCR 反应体系和反应条件，实际操作中应根据模板、引物结构和目的片段大小不同进行相应的改进和优化。
1.PCR反应体系：注：引物浓度以0.1-1.0 μM终浓度作为设定范围的参考。
扩增效率不高时，提高引物的浓度；发生非特异性反应时，可降低引物浓度; DNA模板的量以10-100 ng基因组DNA或1-10 ng cDNA为参照。
2. PCR反应程序：两步法PCR：步骤温度时间预变性95℃10 min变性95℃15 s退火/延伸60℃ 1 min融解曲线分析95℃15 s60℃ 1 min95℃15 s60℃15 s注意：1）本产品所采用的热启动酶须在预变性95℃、10 min条件下实现酶的活化。
一步法实时RTPCR 检测试剂盒（SYBR 一步法荧光定量PCR 试剂盒）(目录号HS0614)产品包装保存条件-20℃避光保存。
产品简介本产品是一步法Real-Time qRT-PCR 专用试剂盒。
所含的SYBR Green I 荧光染料可以与所有的双链DNA 相结合，使该产品可以用于多种不同靶序列的检测而不需合成特异性标记探针。
使用本产品进行Real Time qRT-PCR 反应，逆转录和定量PCR 在同一反应体系中进行，反应过程中无需添加试剂，无需打开管盖，避免了污染的同时提高了实验效率。
本试剂盒中包含全新高效逆转录酶，具有与RNA 高亲和性的特点，能通读GC 含量高、二级结构复杂的RNA 模板；所包含的经化学修饰的热启动DNA 聚合酶最大限度的减少PCR 扩增全程中的非特异性扩增产物的产生，极大提高了荧光定量PCR 反应的精确性；所包含的缓冲系统使以上两种酶同时发挥最大的功效，提高效率。
所含的ROX 染料调节PCR 反应过程中管与管之间的加样误差，适用于以ROX 作为校正染料的所有荧光定量PCR 仪。
北京厚生博泰科技有限公司 Beijing Hooseen Biotech Co., Ltd.产品特点1. 反转录和PCR在一个反应管中完成，方便快捷，最大限度的避免污染。
2. 与RNA具有高亲和性，能通读GC含量高、二级结构复杂的RNA模板。
3. 热启动酶有效抑制非特异性扩增，极大提高了荧光定量PCR反应的精确性。
产品应用基因表达分析；拷贝数分析。
使用方法1. 将RNA 模板、引物、2×UltraSYBR One Step RT-qPCR Buffe（r With ROX）、SuperEnzyme Mix 和RNase-Free Water溶解并置于冰上备用。
2. PCR 反应体系：•注意：通常引物浓度以0.2 uM可以得到较好结果，可以终浓度0.1 ~ 0.5 uM作为设定范围的参考。
