QIAGEN OneStep RT-PCR Kit
用于灵敏、特异性的一步法RT-PCR
Products
QIAGEN OneStep RT-PCR Kit 旨在用于分子生物学应用。该产品不能用于疾病诊断、预防和治疗。
QIAGEN OneStep RT-PCR Kit (100)
Cat. No. / ID: 210212
For 100 x 50 µl reactions: QIAGEN OneStep RT-PCR Enzyme Mix (1 x 200 µl), 5x QIAGEN OneStep RT-PCR Buffer (1 x 1 ml), dNTP Mix (1 x 200 µl, 10 mM each), 5x Q-Solution (1 x 2 ml), RNase-Free Water (2 x 1.9 ml)
QIAGEN OneStep RT-PCR Kit (1000)
Cat. No. / ID: 210215
For 1000 x 50 µl reactions (available in a single tube): QIAGEN OneStep RT-PCR Enzyme Mix (2 x 1 ml), 5x QIAGEN OneStep RT-PCR Buffer (1 x 10 ml), dNTP Mix (2 x 1 ml, 10 mM each), 5x Q-Solution (1 x 10 ml), RNase-Free Water (1 x 40 ml)
QIAGEN OneStep RT-PCR Kit (25)
Cat. No. / ID: 210210
For 25 x 50 µl reactions: QIAGEN OneStep RT-PCR Enzyme Mix (1 x 50 µl), 5x QIAGEN OneStep RT-PCR Buffer (1 x 250 µl), dNTP Mix (1 x 50 µl, 10 mM each), 5x Q-Solution (1 x 400 µl), RNase-Free Water (1 x 1.9 ml)
特点
- 快速方便的单管反应
- 适合各种RNA模板，无需优化反应条件
- 独特的酶组合，确保反应的高特异性和高灵敏度
- 经优化的缓冲体系确保了高效的逆转录和扩增过程
产品详情
QIAGEN One-Step RT-PCR Kit包含Omniscript逆转录酶和Sensiscript逆转录酶的混合物、HotStarTaq DNA Polymerase、QIAGEN OneStep RT-PCR Buffer、dNTP混合液和Q-Solution。Q-Solution是一种新型的辅助剂，有助于实现“困难”模板（如：高GC含量的模板）的高效扩增。简单的单管反应体系构建和优化的试剂盒组分确保了结果成功且灵敏度高。
绩效
QIAGEN OneStep RT-PCR Kit提供方便的规格，可使用RNA模板进行高灵敏度和特异性的RT-PCR。试剂盒中包含优化的组分，可在同一反应混合液中进行逆转录和PCR扩增，完成"一步式"反应。独特的酶组合和专门研发的反应缓冲液确保在一个管中进行高效、高特异性的逆转录和PCR，无需优化（参见" Highly specific RT-PCR using low amounts of template" 和 " Efficient detection of viral RNA"）。试剂盒提供的新型的、双阳离子PCR缓冲液确保在宽退火温度范围内得到高产量的特异性PCR产物（参见" Influence of annealing temperature on specificity"）。使用Q-Solution可提高不理想的RT-PCR的表现，这种独特的辅助剂可帮助逆转录并高效扩增具有高GC含量和复杂二级结构的模板（参见" RT-PCR of GC-rich template"）。
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原理
QIAGEN OneStep RT-PCR Kit可对多种RNA模板进行简单、灵敏的一步式RT-PCR。
QIAGEN OneStep RT-PCR Enzyme Mix含有为逆转录和PCR反应特别设计的酶。独特组合的Omniscript和Sensiscrip逆转录酶对RNA模板有高度亲和性，确保RNA逆转录的灵敏度达到1 pg到2 µg。逆转录完成后，升温至95°C，15分钟，活化HotStarTaq DNA Polymerase同时使逆转录酶失活。这种方法可减少引物二聚体等非特异性扩增产物的生成，并减少背景的影响，确保RT-PCR的高灵敏度和可重复性。
退火温度范围宽
引物的最佳退火温度取决于碱基对的组成（如A、T、G、C核苷酸的比例）、引物的浓度和反应的离子环境。QIAGEN PCR Buffers含有K+和NH4+，使用该缓冲液可在很宽的温度范围内增加特异性PCR产物的产量（参见" Increased specific primer annealing"）。其原理是破坏非特异性结合的引物对，提供更加可靠的反应环境，减少繁琐的温度优化过程。相反，如使用仅含K+的PCR或一步式RT-PCR缓冲液，PCR最佳退火温度的范围较窄，可靠性较差。
QIAGEN OneStep RT-PCR Buffer专为提高逆转录和PCR扩增的效率而设计。该缓冲液含有新型辅助剂，可防止逆转录酶对PCR扩增的抑制，而这是一步式RT-PCR中常见的问题。该缓冲液确保在较大的温度范围和Mg2+浓度范围内，引物与模板特异性结合，可对多种RNA模板进行高效的RT-PCR扩增。QIAGEN OneStep RT-PCR Kit含有Q-Solution，这是可修饰核酸熔解行为的新型辅助剂。Q-Solution可促进高GC含量或含较多二级结构的模板的逆转录和扩增。 使用Q-Solution可简化复杂模板RT-PCR的优化过程。QIAGEN OneStep RT-PCR Kit含有您所需的所有试剂，可进行快速、简便的RT-PCR，用于多种灵敏的应用（见表）。
| QIAGEN OneStep RT-PCR Kit 组分 | 特点 |
|---|---|
| HotStarTaq DNA Polymerase | 高度特异性产物 |
| Sensiscript and Omniscript Reverse Transcriptases | RNA初始量的范围大 (1 pg–2 µg) 高灵敏度 |
| OneStep RT-PCR Buffer | 无需优化 反转录酶不会抑制PCR 抑制RNases |
| Q-Solution | 促进GC含量高的模板的扩增 |
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程序
使用QIAGEN OneStep RT-PCR Kit可快速、简单的进行RT-PCR反应体系构建。无论是何种应用，即无论是病毒检测、分子诊断研究还是基因表达，您只需将所有试剂在一个离心管中混合即可开始PCR扩增（参见" OneStep RT-PCR procedure"）。反应混合液含有逆转录和PCR反应所需的所有试剂，无需添加其他试剂即可开始反应（参见下表）。
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应用
QIAGEN OneStep RT-PCR Kit适用于RT-PCR分析，包括：
- 病毒检测
- 单细胞RT-PCR
- 基因表达分析
辅助数据和图表
OneStep RT-PCR procedure.
The QIAGEN OneStep RT-PCR Kit allows fast and easy RT-PCR setup. Whatever the application — virus detection, molecular diagnostics research, or gene expression analysis— just mix all components together in one tube and start the thermal-cycler program. The reaction mixture contains all of the reagents required for both reverse transcription and PCR; nothing needs to be added once the reaction has been started.
Specifications
| Features | Specifications |
|---|---|
| Applications | Gene-expression analysis, virus detection |
| Mastermix | No |
| Enzyme activity | Reverse transcription, 5' -> 3' exonuclease activity |
| Reaction type | One-step RT-PCR |
| Real-time or endpoint | Endpoint |
| Sample/target type | RNA template |
| Single or multiplex | Single |
| With/without hotstart | With hotstart |
资源
补充实验方案 (6)
产品介绍与指南 (3)
试剂盒操作手册 (1)
快速启动实验方案 (1)
安全数据表 (1)
技术资讯 (1)
Safety Data Sheets (1)
Certificates of Analysis (1)
Publications
Up-regulation of cyclooxygenase-2 expression is involved in R(+)-methanandamide-induced apoptotic death of human neuroglioma cells.
Mol Pharmacol; 2004; 66 (6):1643-51 2004 Sep 10 PMID:15361550
Drosophila crinkled, mutations of which disrupt morphogenesis and cause lethality, encodes fly myosin VIIA.
Genetics; 2004; 168 (3):1337-52 2004 Nov PMID:15579689
Instability of the restriction fragment length polymorphism pattern of open reading frame 5 of porcine reproductive and respiratory syndrome virus during sequential pig-to-pig passages.
J Clin Microbiol; 2004; 42 (10):4462-7 2004 Oct PMID:15472294
Expression of growth differentiation factor 9 messenger RNA in porcine growing and preovulatory ovarian follicles.
Biol Reprod; 2004; 71 (4):1290-5 2004 Jun 9 PMID:15189836
Effect of vaccine use in the evolution of Mexican lineage H5N2 avian influenza virus.
J Virol; 2004; 78 (15):8372-81 2004 Aug PMID:15254209
FAQ
Are there specific recommendations for performing RT-PCR on RNA isolated from paraffin-embedded samples?
Do I need to use an RNase inhibitor in my RT reaction?
Is mRNA isolation necessary for sensitive RT-PCR?
Does QIAGEN sell Q-Solution separately?
Do you have a protocol for polyacrylamide gel analysis of oligonucleotides?
Does the 5x OneStep RT-PCR Buffer contain BSA?
Can I shorten the activation time for the HotStarTaq DNA Polymerase?