QuantiFast SYBR® Green PCR Kit

For fast, real-time PCR and two-step qRT-PCR using SYBR Green

  • Specific and sensitive detection of even low-copy targets
  • Optimized master mix for reliable results without optimization
  • 5-minute enzyme activation and fast cycling conditions
  • Accurate detection of a wide range of template amounts
  • Universal protocol for all standard and fast cyclers
The QuantiFast SYBR Green PCR Kit delivers fast and specific quantification of gDNA or cDNA targets by real-time PCR or two-step RT-PCR using SYBR Green I detection. Q-bond technology and an optimized, ready-to-use 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. For convenience, the master mix can be stored at 2–8°C.
产品 货号 目录价:
QuantiFast SYBR Green PCR Kit (4000)
For 4000 x 25 µl reactions: 2 x 25 ml 2x QuantiFast SYBR Green PCR Master Mix (contains ROX dye), 20 ml RNase-Free Water
204057
询价

QuantiFast SYBR® Green PCR Kit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。


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PCR反应时间极大缩短。|能够分辨拷贝数的细微差别。|更快获得结果,且灵敏性高。|灵敏的两步法RT-PCR。|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 SYBR® Green PCR 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,无熔曲线分析;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 SYBR® Green PCR Kit检测男性供体基因组DNA中的Y染色体特异性的单拷贝基因SRY。[A] 可明显区分出1000至1.5拷贝的曲线。[B] 拷贝数 (log) 与CT值曲线显示了高线性度。|在Applied Biosystems 7500 Fast System上利用两步法real-time RT-PCR分析人白细胞中MYC (一种原癌基因) 的表达。使用10倍稀释的cDNA (10 ng至10 pg) 进行两次重复反应。[A] 在快速循环模式下使用QuantiFast Kit的CT值与 [B] 在标准循环模式下使用QuantiTect Kit的CT值相当。而使用Supplier AII的标准循环试剂盒,不仅是在 [C] 快速循环模式下,而且在 [D] 推荐的标准循环模式下,亦只能获得较差的CT值。|使用iCycler iQ和 [A] QuantiFast SYBR® Green PCR Kit或 [B] Supplier BV的试剂盒,分析人白细胞中MYC (一种原癌基因) 的表达。使用10倍稀释的cDNA (100 ng至0.1 ng) 进行两次重复反应。参照快速循环实验方案,使用Supplier BV的仪器专用试剂盒。QuantiFast Kit的CT值较低,说明灵敏度更高。|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

The QuantiFast SYBR Green PCR Kit delivers highly specific and sensitive results, outperforming other real-time PCR kits used in fast cycling mode (see figure "Sensitive two-step RT-PCR"). PCR run times are reduced by up to 60% (see figure "Significantly reduced PCR times"), allowing you to get results faster without compromising 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. 

The QuantiFast SYBR Green PCR Kit enables accurate quantification of targets over several log dilutions of template. Even small differences in the amount of low-copy targets can be clearly distinguished (see figure "Resolution of small differences in copy number").

原理

The QuantiFast SYBR Green PCR Kit delivers highly sensitive and specific results over a wide dynamic range on both standard and fast cyclers. The fluorescent dye SYBR Green I in the master mix enables the analysis of many different targets without having to synthesize target-specific labeled probes. A specially developed fast PCR buffer contains the novel PCR additive Q-Bond, which significantly reduces denaturation, annealing, and extension times (see figure "Fast primer annealing"). A balanced combination of K+ and NH4+ ions in the PCR buffer promotes specific primer annealing and enables high 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 SYBR Green 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 SYBR Green 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
SYBR Green I dye Yields a strong fluorescent signal upon binding to double-stranded DNA Highly sensitive amplification
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 a dNTP mix (dATP, dCTP, dGTP, dTTP).
操作流程

The QuantiFast SYBR Green PCR Kit is a ready-to-use master mix that eliminates the need for optimization of reaction and cycling conditions. Simply add primers and DNA template to the ready-to-use PCR master mix, and start the reaction. Follow the protocol in the handbook to get fast and reliable results on any real-time cycler.

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.

We also recommend QuantiTect Primer Assays for gene expression analysis using SYBR Green. QuantiTect Primer Assays are bioinformatically validated primer sets for any gene from human, mouse, rat, and many other species. Assays can be easily ordered online at the GeneGlobe Web portal.

 

应用

The QuantiFast SYBR Green PCR Kit is for use in gene expression analysis of cDNA targets and quantitative gDNA analysis. QuantiFast SYBR Green PCR Kits are compatible with all available real-time cyclers, including 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 SYBR Green PCR Kit, which has been specially developed for fast cycling on these instruments.

特点
参数
应用 SYBR Green-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 SYBR Green I
热循环仪 Applied Biosystems, Bio-Rad, Cepheid, QIAGEN, Eppendorf, Roche, and Agilent
有/无ROX With ROX
有/无热启动酶 With hotstart

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试剂盒操作手册
2
For fast, quantitative, real-time PCR and two-step RT-PCR using SYBR Green I
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For genomewide, ready-to-use real-time RT-PCR assays using SYBR Green detection
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安全数据表
1
Download Safety Data Sheets for QIAGEN product components.
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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 SYBR® Green PCR 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,无熔曲线分析;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。
Resolution of small differences in copy number on the Mastercycler ep realplex.
能够分辨拷贝数的细微差别。
在Mastercycler ep realplex上使用QuantiFast SYBR® Green PCR Kit检测男性供体基因组DNA中的Y染色体特异性的单拷贝基因SRY。[A] 可明显区分出1000至1.5拷贝的曲线。[B] 拷贝数 (log) 与CT值曲线显示了高线性度。
Faster results without compromising sensitivity on the Applied Biosystems 7500 Fast System.
更快获得结果,且灵敏性高。
在Applied Biosystems 7500 Fast System上利用两步法real-time RT-PCR分析人白细胞中MYC (一种原癌基因) 的表达。使用10倍稀释的cDNA (10 ng至10 pg) 进行两次重复反应。[A] 在快速循环模式下使用QuantiFast Kit的CT值与 [B] 在标准循环模式下使用QuantiTect Kit的CT值相当。而使用Supplier AII的标准循环试剂盒,不仅是在 [C] 快速循环模式下,而且在 [D] 推荐的标准循环模式下,亦只能获得较差的CT值。
Senstivite two-step RT-PCR on the iCycler iQ.
灵敏的两步法RT-PCR。
使用iCycler iQ和 [A] QuantiFast SYBR® Green PCR Kit或 [B] Supplier BV的试剂盒,分析人白细胞中MYC (一种原癌基因) 的表达。使用10倍稀释的cDNA (100 ng至0.1 ng) 进行两次重复反应。参照快速循环实验方案,使用Supplier BV的仪器专用试剂盒。QuantiFast Kit的CT值较低,说明灵敏度更高。
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。