QuantiFast Probe PCR Kits

For fast, real-time PCR and two-step qRT-PCR using sequence-specific probes

  • Sensitive detection of even low copy targets
  • Accurate detection of a wide range of template amounts
  • Faster results with a time savings of up to 60%
  • Optimized, ready-to-use master mix for reliable results without optimization
  • One cycling protocol for all standard and fast cyclers
QuantiFast Probe PCR Kits deliver fast and sensitive quantification of gDNA and cDNA targets by real-time PCR and two-step RT-PCR using sequence-specific probes. Q-bond technology and an optimized master mix enable shorter real-time 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. Two kit formats are available: the QuantiFast Probe PCR Kit for cyclers that require ROX dye for fluorescence normalization, and the QuantiFast Probe PCR +ROX Vial Kit for all other cyclers. For convenience, the master mix can be stored at 2–8°C.
产品 Product no. 货号 目录价:
 
 
Show details
    varies
Can't order online?
To place an order via phone, email or for requesting a quote, please provide the product’s name, number and catalog number.
产品 货号 目录价:
QuantiFast Probe PCR Kit (400)
For 400 x 25 µl reactions: 3 x 1.7 ml 2x QuantiFast Probe PCR Master Mix (contains ROX dye), 2 x 2 ml RNase-Free Water
204254
7.020,00 kr
加入购物车
QuantiFast Probe PCR Kit (2000)
For 2000 x 25 µl reactions: 25 ml 2x QuantiFast Probe PCR Master Mix (contains ROX dye), 20 ml RNase-Free Water
204256
27.685,00 kr
加入购物车
QuantiFast Probe PCR Kit (4000)
For 4000 x 25 µl reactions: 2 x 25 ml 2x QuantiFast Probe PCR Master Mix (contains ROX dye), 20 ml RNase-Free Water
204257
38.780,00 kr
加入购物车
QuantiFast Probe PCR +ROX Vial Kit (400)
For 400 x 25 µl reactions: 3 x 1.7 ml 2x QuantiFast Probe PCR Master Mix (without ROX dye), 210 µl ROX Dye Solution, 2 x 2 ml RNase-Free Water
204354
7.020,00 kr
加入购物车
QuantiFast Probe PCR +ROX Vial Kit (2000)
For 2000 x 25 µl reactions: 25 ml 2x QuantiFast Probe PCR Master Mix (without ROX dye), 1.05 ml ROX Dye Solution, 20 ml RNase-Free Water
204356
27.685,00 kr
加入购物车

QuantiFast Probe 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。


See trademarks.

  • Main Image Navi
PCR反应时间极大缩短。|线性范围广,灵敏度高。|更快获得结果,且保持高灵敏性。|灵敏的两步法RT-PCR。|检测具有宽泛的动态范围。|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进行两步法real-time RT-PCR,可缩短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Mastercycler ep realplex上使用QuantiFast Probe PCR +ROX Vial Kit进行两步法PCR,可实现从低模板量至高模板量的准确基因表达分析,PCR效率为93%。使用10倍稀释的人白细胞cDNA和采用Primer Express设计的针对泛素 (一种调节蛋白) 的TaqMan Assay进行两次重复反应 (25 μl体积)。NTC:无模板对照。|在ABI PRISM 7900上使用10倍稀释的人白细胞cDNA (100 ng至0.01 ng) 和针对IL1RN (一种细胞因子) 的TaqMan Gene Expression Assay进行两次重复反应。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。|在ABI PRISM 7900上使用10倍稀释的人白细胞cDNA (100 ng至0.01 ng) 和采用Primer Express设计的针对IL8 (一种趋化因子) 的TaqMan Assay进行三次重复反应。[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0.01 ng cDNA的转录本定量。|在LightCycler 2.0上使用Supplier R提供的靶向b-2微球蛋白的一组引物和FRET探针,进行双重两步法RT-PCR反应。QuantiFast Kit的扩增曲线显示其具有较宽的动态范围。|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 PCR Kits deliver highly sensitive results, outperforming other real-time PCR kits (see table "Sensitive two-step RT-PCR on the iCycler iQ" and figure and Sensitive two-step RT-PCR). 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 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.

Sensitive two-step RT-PCR on the iCycler iQ
Mean CT value
Human leukocyte cDNA (ng) QuantiFast Probe PCR +ROX Vial Kit Kit from Supplier BV
100 29.13 31.13
10 32.50 34.43
1 36.15 40.00
No template control 40.00 40.00
Triplicate reactions were run using a Primer Express designed TaqMan® assay for IL8 (a chemokine). The QuantiFast Kit provided lower CT values than the kit from Supplier BV (which was used according to a fast-cycling protocol), allowing transcript quantification from just 1 ng cDNA.

QuantiFast Probe PCR Kits allow accurate quantification over a wide dynamic range (see figure "Wide dynamic range and high sensitivity"). Template dilutions of up to 7 logs can be reliably detected (see table "Detection over 7 logs of template in two-step RT-PCR on the LightCycler 2.0 " and figure "Detection over 7 logs of template").

Detection over 7 logs of template in two-step RT-PCR on the LightCycler 2.0
Mean CT value
Human leukocyte cDNA QuantiFast Probe PCR +ROX Vial Kit Kit from Supplier R
100 ng 11.99 14.56
10 ng 15.54 17.99
1 ng 19.22 22.16
100 pg 22.63 25.14
10 pg 26.43 30.15
1 pg 30.02 34.43
100 fg 33.42 37.10
No template control 40 40.00
PCR efficiency 90% 80.1%
Duplicate reactions were run using a set of primers and FRET probes for β-2 microglobulin from supplier R. The QuantiFast Kit provided lower CT values and higher PCR efficiency that the kit from Supplier R (which was used according to the fast cycling protocol ).
原理

QuantiFast Probe 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. A specially developed, fast PCR buffer contains the novel additive Q-Bond, which significantly reduces denaturation, annealing, and extension times (see figure "Fast primer annealing"). The buffer also contains a balanced combination of K+ and NH4+ ions, which promote specific primer annealing, enabling high PCR specificity and sensitivity (see figure "Specific primer annealing"). In addition, HotStarTaq Plus DNA Polymerase provides a stringent hot start, preventing the formation of nonspecific products.

Components of 2x QuantiFast Probe PCR Kit*
Component Features and benefits Benefits
HotStarTaq Plus DNA Polymerase
3 min activation at 95ºC
Set up of qPCR reactions at room temperature
QuantiFast Probe 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
* 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 PCR Kits contain ready-to-use master mixes that eliminate the need for optimization of reaction and cycling conditions. Simply add template DNA, 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 PCR Kit
ROX dyeKit Compatible cyclers
Supplied in master mix QuantiFast Probe PCR Kit All cyclers from Applied Biosystems except Applied Biosystems 7500
Supplied in a separate tube QuantiFast Probe PCR +ROX Vial Kit Applied Biosystems 7500, and cyclers from Agilent, Bio-Rad, Cepheid, QIAGEN, Eppendorf, Roche, and other suppliers
.

For optimal results in real-time two-step RT-PCR, we recommend synthesizing cDNA using the QuantiTect Reverse Transcription Kit. The kit provides fast cDNA synthesis in just 20 minutes with integrated removal of genomic DNA contamination. 

QuantiFast Probe Assays are predesigned, genomewide assays that use hydrolysis, probe-based detection. They are delivered with the QuantiFast Probe PCR Kit for guaranteed results in singleplex, two-step qRT-PCR.

应用

QuantiFast Probe PCR Kits can be used for probe-based gene expression analysis of cDNA targets and quantitative gDNA analysis 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 PCR Kit, which has been specially developed for fast cycling on these instruments.

特点
参数
应用 Probe-based, real-time PCR, two-step RT-PCR
反应类型 Real-time and two-step RT-PCR
real-time或终点法PCR Real-time
样本/目标类型 cDNA, DNA
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. Roche LightCycler®, Corbett Rotor-Gene, Applied Biosystems)
有/无ROX Available with ROX in master mix and with ROX as separate vial

您无权限查看此资源

常见问题
37
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 -1432
浏览
FAQ ID -1433
浏览
FAQ ID -1435
浏览
FAQ ID -1436
浏览
FAQ ID -1437
浏览
FAQ ID -1438
浏览
FAQ ID -1440
浏览
FAQ ID -1441
浏览
FAQ ID -1442
浏览
FAQ ID -1446
浏览
FAQ ID -1447
浏览
FAQ ID -1449
浏览
FAQ ID -1554
浏览
FAQ ID -1555
浏览
FAQ ID -1592
浏览
FAQ ID -1697
浏览
FAQ ID -1702
浏览
FAQ ID -2119
浏览
FAQ ID -2125
浏览
FAQ ID -2362
浏览
FAQ ID -2371
浏览
FAQ ID -2373
浏览
FAQ ID -2651
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
FAQ ID -1452
浏览
试剂盒操作手册
1
For fast, quantitative, real-time PCR and two-step RT-PCR using sequence-specific probes
显示更多
图片
Significantly reduced PCR times
PCR反应时间极大缩短。
在标准PCR循环仪上使用QuantiFast Probe Kit进行两步法real-time RT-PCR,可缩短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。
Wide dynamic range and high sensitivity in two-step RT-PCR on the Mastercycler ep realplex.
线性范围广,灵敏度高。
在Mastercycler ep realplex上使用QuantiFast Probe PCR +ROX Vial Kit进行两步法PCR,可实现从低模板量至高模板量的准确基因表达分析,PCR效率为93%。使用10倍稀释的人白细胞cDNA和采用Primer Express设计的针对泛素 (一种调节蛋白) 的TaqMan Assay进行两次重复反应 (25 μl体积)。NTC:无模板对照。
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) 和针对IL1RN (一种细胞因子) 的TaqMan Gene Expression Assay进行两次重复反应。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。
Sensitive two-step RT-PCR on the ABI PRISM 7900.
灵敏的两步法RT-PCR。
在ABI PRISM 7900上使用10倍稀释的人白细胞cDNA (100 ng至0.01 ng) 和采用Primer Express设计的针对IL8 (一种趋化因子) 的TaqMan Assay进行三次重复反应。[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0.01 ng cDNA的转录本定量。
Detection over 7 logs of template in two-step RT-PCR on the LightCycler 2.0.
检测具有宽泛的动态范围。
在LightCycler 2.0上使用Supplier R提供的靶向b-2微球蛋白的一组引物和FRET探针,进行双重两步法RT-PCR反应。QuantiFast Kit的扩增曲线显示其具有较宽的动态范围。
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。