QuantiFast Probe RT-PCR Kits

For fast, one-step qRT-PCR using sequence-specific probes for gene expression analysis

  • Faster results with time savings of up to 60%
  • Sensitive detection of even low copy numbers
  • Accurate detection of a wide range of template amounts
  • Optimized, ready-to-use master mix for fast cycling
  • One cycling protocol for all standard and fast cyclers
QuantiFast Probe RT-PCR Kits deliver fast and sensitive real-time one-step RT-PCR quantification of RNA targets using probe-based detection. Q-bond technology and an optimized master mix enable shorter real-time RT-PCR run times, not only on fast cyclers with short ramping times, but also on standard cyclers. The combination of a hot start and a unique PCR buffer system in the ready-to-use master mix ensures highly sensitive qPCR on any real-time cycler without the need for optimization. The kits are also supplied with an optimized RT mix for efficient cDNA synthesis in only 10 minutes. Two kit formats are available: the QuantiFast Probe RT-PCR Kit for cyclers that require ROX dye for fluorescence normalization, and the QuantiFast Probe RT-PCR +ROX Vial Kit for all other cyclers. For convenience, the master mix can be stored at 2–8°C.
产品 货号 目录价:
QuantiFast Probe RT-PCR Kit (400)
For 400 x 25 µl reactions: 3 x 1.7 ml 2x QuantiFast Probe RT-PCR Master Mix (contains ROX dye), 100 µl QuantiFast RT Mix, 2 x 2 ml RNase-Free Water
204454
询价
QuantiFast Probe RT-PCR Kit (2000)
For 2000 x 25 µl reactions: 25 ml 2x QuantiFast Probe RT-PCR Master Mix (contains ROX dye), 0.5 ml QuantiFast RT Mix, 20 ml RNase-Free Water
204456
询价
QuantiFast Probe RT-PCR +ROX Vial Kit (400)
For 400 x 25 µl reactions: 3 x 1.7 ml 2x QuantiFast Probe RT-PCR Master Mix (without ROX dye), 210 µl ROX Dye Solution, 100 µl QuantiFast RT Mix, 2 ml RNase-Free Water
204554
询价
QuantiFast Probe RT-PCR +ROX Vial Kit (2000)
For 2000 x 25 µl reactions: 25 ml 2x QuantiFast Probe RT-PCR Master Mix (without ROX dye), 1.05 ml ROX Dye Solution, 0.5 ml QuantiFast RT Mix, 20 ml RNase-Free Water
204556
询价

QuantiFast Probe RT-PCR Kits适用于分子生物学应用。该产品不适用于疾病的诊断、预防或治疗。


  • Main Image Navi
RT-PCR反应时间明显缩短。|灵敏的一步法RT-PCR。|动态范围宽。|更快获得结果,且保持高灵敏性。|动态范围宽,灵敏度高。|退火时引物特异性结合。|退火时引物快速结合。|
在标准PCR循环仪上使用QuantiFast Probe Kit进行一步法real-time RT-PCR,可缩短RT-PCR总运行时间达60% (进行40个PCR循环;与标准的QIAGEN real-time PCR Kit相比较)。L:LightCycler 2.0;A1:ABI PRISM 7900;A2:Applied Biosystems 7500;A3:ABI PRISM 7000。|在Applied Biosystems 7500 Fast System上使用10倍稀释的人白细胞RNA (100 ng至0.1 ng) 和采用Primer Express设计的针对IL12RBI (一种I型跨膜蛋白) 的TaqMan Assay进行两次重复反应。[A] QuantiFast Kit的灵敏度高于 [B] Supplier I的试剂盒 (参照标准循环实验方案使用),具有更低的CT值,可实现低至0.1 ng RNA的转录本定量。|使用Primer Express设计的TaqMan Assay对人白细胞中的泛素表达进行分析。在ABI PRISM 7900上进行双重两步法和一步法qPCR反应。[A] 使用QuantiFast Probe PCR Kit分析10倍稀释的cDNA (100 ng至10 pg)。[B] 使用QuantiFast Probe RT-PCR Kit分析10倍稀释的RNA (100 ng至10 pg)。|在ABI PRISM 7900上使用10倍稀释的人白细胞cDNA (100 ng至0.01 ng) 和28S rRNA的TaqMan Gene Expression Assay进行反应。QuantiFast Kit的灵敏度高于Supplier AII的标准循环试剂盒 (参照标准循环实验方案使用),具有更低的CT值,可实现低至10 pg cDNA的转录本定量。|使用10倍稀释的人白细胞RNA和针对β-actin的TaqMan Assay重复三次反应。QuantiFast Probe RT-PCR Kit提供了100 ng至1 pg RNA模板范围的准确基因表达分析。|退火时,缓冲液中浓度平衡的KCl和(NH4)2SO4促进引物和探针特异性与模板结合。K+结合到双链DNA上的磷酸基团,稳定引物和探针。NH4+则破坏弱的错配碱基之间的氢键。|[A] QuantiFast Buffer中的Q-Bond能使DNA聚合酶和引物结合为单一复合物,将退火时间缩短至几秒。此外,独特的缓冲液组分可支持DNA的熔解,缩短变性和延伸时间。[B] 若无Q-Bond,引物和聚合酶会相继与模板结合,退火时间较长。|
性能

QuantiFast Probe RT-PCR Kits deliver highly sensitive results, outperforming other real-time RT-PCR kits (see figure "Sensitive One-step RT-PCR"). RT-PCR run times are reduced by up to 60% (see figure "Significantly reduced PCR times"), allowing you to achieve fast PCR results without compromising on RT-PCR performance (see figure "Faster results without compromising sensitivity"). You can also greatly increase your sample throughput or efficiently share a cycler with other users. With QuantiFast Probe RT-PCR Kits, you follow the same procedure whether you work with a standard or fast cycler.

QuantiFast Probe RT-PCR Kits allow accurate quantification over a wide dynamic range (see figure "Comparable dynamic range"). Template dilutions of up to 6 logs can be reliably detected (see figure "Wide dynamic range and high sensitivity").

原理

QuantiFast Probe RT-PCR Kits deliver highly sensitive and rapid results over a wide dynamic range on both standard and fast cyclers. The kits are designed for use with all types of sequence-specific probes, including hydrolysis probe detection (e.g., TaqMan and other dual-labeled probes) and FRET probes. The optimized QuantiFast RT Mix enables cDNA synthesis in just 10 minutes and a specially developed fast RT-PCR buffer contains the novel additive Q-Bond, which significantly reduces denaturation, annealing, and extension times (see figure "Fast primer annealing"). The RT-PCR buffer also contains a balanced combination of K+ and NH4+ ions, which promote specific primer annealing and enable high RT-PCR specificity and sensitivity (see figure "Specific primer annealing"). In addition, HotStarTaq Plus DNA Polymerase requires only 5 minutes at 95°C for activation and provides a stringent hot start, preventing the formation of nonspecific products.  

Components of 2x QuantiFast Probe RT-PCR Kit*
Component Features and benefits Benefits
HotStarTaq Plus DNA Polymerase 5 min activation at 95ºC Set up of qPCR reactions at room temperature
QuantiFast Probe RT-PCR Buffer Balanced combination of NH4+ and K+ ions Specific primer annealing ensures reliable PCR results
Unique Q-Bond additive Faster PCR run times, enabling faster results and more reactions per day
ROX dye Normalizes fluorescent signals on Applied Biosystems and, optionally, Agilent instruments Precise quantification on cyclers that require ROX dye. Does not interfere with PCR on any real-time cycler
QuantiFast RT Mix Separate solution added during PCR setup Fast cDNA synthesis in just 10 minutes
* Also contains dNTP mix (dATP, dCTP, dGTP, and dTTP).
ROX dye is either present in the master mix or as a separate solution.
操作流程

QuantiFast Probe RT-PCR Kits contain ready-to-use master mixes that eliminate the need for optimization of reaction and cycling conditions. Simply add template RNA, primers, and probe to the master mix and follow the protocol in the handbook to get fast and reliable results on any real-time cycler. Kits are available with or without ROX passive reference dye in the master mix, enabling use on virtually any real-time cycler (see table).  Due to the optimized ROX concentrations, detection of even low copy numbers is achieved through automatic data analysis.

Choosing the right QuantiFast Probe RT-PCR Kit
ROX dyeKit Compatible cyclers
Supplied in master mix QuantiFast Probe RT-PCR Kit All cyclers from Applied Biosystems except Applied Biosystems 7500
Supplied in separate tube QuantiFast Probe RT-PCR +ROX Vial Kit Applied Biosystems 7500 and cyclers from
Bio-Rad, Cepheid, Eppendorf, QIAGEN, Roche, Agilent, and other suppliers
应用

QuantiFast Probe RT-PCR Kits can be used for probe-based gene expression analysis of RNA targets on any real-time cycler. This includes instruments from Applied Biosystems, Bio-Rad, Cepheid, Eppendorf, Roche, and Agilent. For the Rotor-Gene Q and other Rotor-Gene cyclers, we recommend using the Rotor-Gene Probe RT-PCR Kit, which has been specially developed for fast cycling on these instruments.

特点
参数
应用 Probe-based, real-time RT-PCR
反应类型 Real-time one-step RT-PCR
real-time或终点法PCR Real-time
样本/目标类型 RNA
单一或多重 Single
SYBR Green I或序列特异性探针 Sequence-specific probes
热循环仪 All real-time cyclers (e.g. LC, RG, ABI)
有/无ROX Available with ROX in master mix and with ROX as a separate vial

您无权限查看此资源

常见问题
27
FAQ ID -1085
浏览
FAQ ID -1425
浏览
FAQ ID -1427
浏览
FAQ ID -1428
浏览
FAQ ID -1429
浏览
FAQ ID -1430
浏览
FAQ ID -1431
浏览
FAQ ID -1437
浏览
FAQ ID -1440
浏览
FAQ ID -1441
浏览
FAQ ID -1442
浏览
FAQ ID -1446
浏览
FAQ ID -1451
浏览
FAQ ID -1695
浏览
FAQ ID -1697
浏览
FAQ ID -1702
浏览
FAQ ID -2105
浏览
FAQ ID -2119
浏览
FAQ ID -2358
浏览
FAQ ID -2371
浏览
FAQ ID -2655
浏览
FAQ ID -2682
浏览
FAQ ID -539
浏览
FAQ ID -589
浏览
FAQ ID -729
浏览
FAQ ID-751
浏览
FAQ ID -784
浏览
试剂盒操作手册
1
QuantiFast Probe RT-PCR Kit
QuantiFast Probe RT-PCR +ROX Vial Kit For fast, quantitative, real-time, one-step RT-PCR using sequence-specific probes
显示更多
安全数据表
1
Download Safety Data Sheets for QIAGEN product components.
浏览
图片
Significantly reduced PCR times
RT-PCR反应时间明显缩短。
在标准PCR循环仪上使用QuantiFast Probe Kit进行一步法real-time RT-PCR,可缩短RT-PCR总运行时间达60% (进行40个PCR循环;与标准的QIAGEN real-time PCR Kit相比较)。L:LightCycler 2.0;A1:ABI PRISM 7900;A2:Applied Biosystems 7500;A3:ABI PRISM 7000。
Sensitive one-step RT-PCR on the Applied Biosystems 7500 Fast System.
灵敏的一步法RT-PCR。
在Applied Biosystems 7500 Fast System上使用10倍稀释的人白细胞RNA (100 ng至0.1 ng) 和采用Primer Express设计的针对IL12RBI (一种I型跨膜蛋白) 的TaqMan Assay进行两次重复反应。[A] QuantiFast Kit的灵敏度高于 [B] Supplier I的试剂盒 (参照标准循环实验方案使用),具有更低的CT值,可实现低至0.1 ng RNA的转录本定量。
Comparable dynamic range in two-step and one-step RT-PCR on the ABI PRISM 7900.
动态范围宽。
使用Primer Express设计的TaqMan Assay对人白细胞中的泛素表达进行分析。在ABI PRISM 7900上进行双重两步法和一步法qPCR反应。[A] 使用QuantiFast Probe PCR Kit分析10倍稀释的cDNA (100 ng至10 pg)。[B] 使用QuantiFast Probe RT-PCR Kit分析10倍稀释的RNA (100 ng至10 pg)。
Faster results without compromising sensitivity on the ABI PRISM 7900
更快获得结果,且保持高灵敏性。
在ABI PRISM 7900上使用10倍稀释的人白细胞cDNA (100 ng至0.01 ng) 和28S rRNA的TaqMan Gene Expression Assay进行反应。QuantiFast Kit的灵敏度高于Supplier AII的标准循环试剂盒 (参照标准循环实验方案使用),具有更低的CT值,可实现低至10 pg cDNA的转录本定量。
Wide Dynamic Range and High Sensitivity in One-Step RT-PCR on the ABI PRISM 7900
动态范围宽,灵敏度高。
使用10倍稀释的人白细胞RNA和针对β-actin的TaqMan Assay重复三次反应。QuantiFast Probe RT-PCR Kit提供了100 ng至1 pg RNA模板范围的准确基因表达分析。
Specific primer annealing using a balanced combination of K+ and NH4+ ions
退火时引物特异性结合。
退火时,缓冲液中浓度平衡的KCl和(NH4)2SO4促进引物和探针特异性与模板结合。K+结合到双链DNA上的磷酸基团,稳定引物和探针。NH4+则破坏弱的错配碱基之间的氢键。
Fast primer annealing with Q-Bond
退火时引物快速结合。
[A] QuantiFast Buffer中的Q-Bond能使DNA聚合酶和引物结合为单一复合物,将退火时间缩短至几秒。此外,独特的缓冲液组分可支持DNA的熔解,缩短变性和延伸时间。[B] 若无Q-Bond,引物和聚合酶会相继与模板结合,退火时间较长。