用于在Rotor-Gene PCR仪上使用序列特异性探针超快速进行一步法qRT-PCR

  • Rotor-Gene PCR仪上超快速、可靠的结果
  • 灵敏检测低拷贝数基因
  • 大范围模板量内确保准确的检测
  • 专用的即用型预混液确保快速循环

Rotor-Gene Probe RT-PCR Kit专用于Rotor-Gene Q实时荧光定量PCR分析仪和其他Rotor-Gene PCR仪,使用序列特异性探针采用定量一步法RT-PCR可超快速、高度灵敏的定量RNA靶分子。特殊优化的预混液配合独特的Rotor-Gene PCR仪,表现卓越。预混液可贮存在2–8°C,方便使用。

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Rotor-Gene Probe RT-PCR Kit (400)
For 400 x 25 µl reactions: 3 x 1.7 ml 2x Rotor-Gene Probe RT-PCR Master Mix, 100 µl Rotor-Gene RT Mix, 2 x 2 ml RNase-Free Water
204574
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The Rotor-Gene Probe RT-PCR Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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Sensitive detection using a sequence-specific probe.|Specific primer annealing.|Fast primer annealing.|
Tenfold dilutions of human leukocyte RNA (100 ng to 1 ng) were used as template in real-time one-step RT-PCR. Triplicate reactions were run using a TaqMan Gene Expression Assay for IL12RB1 (interleukin 12 receptor, beta 1). Greater sensitivity (i.e., lower CT values) was achieved with [A] the Rotor-Gene Q and Rotor-Gene Probe RT-PCR Kit than with [B] an instrument and kit from Supplier AII.|Cations in the Rotor-Gene Q PCR buffer increase specific primer annealing. K+ binds to phosphate groups on double-stranded DNA, stabilizing primer annealing. NH4+ destabilizes weak hydrogen bonds between mismatched bases.|[A] Q-Bond in Rotor-Gene Probe RT-PCR Master Mix increases the affinity of DNA polymerase for short single-stranded DNA, reducing primer annealing time to a few seconds. In addition, the unique buffer composition supports the melting of DNA, reducing denaturation and extension times. [B] Without Q-Bond, the primer and polymerase bind sequentially to the template, increasing primer annealing time.|
Performance
Rotor-Gene Probe RT-PCR Kit对于低丰度的转录子也可进行可靠的定量分析(参见"Sensitive detection using a sequence-specific probe")。
Principle

Rotor-Gene Probe RT-PCR Kit专用于配合Rotor-Gene Q实时荧光定量PCR分析仪,进行高度可靠的RNA模板定量分析,无需优化反应或扩增条件。该试剂盒专用于水解型探针(如TaqMan探针)。反应以RNA为模板,在同一个反应管中进行逆转录和PCR。由于无需将RT反应产物转移到另一个PCR反应管中,real-time RT-PCR是流水线式的,支持高通量分析。

含有K+和NH4+平衡组合的预混液促进高特异性引物退火 ,确保PCR的高度灵敏和特异性扩增(参见"Specific primer annealing")。新颖的PCR添加剂Q-Bond缩短了循环时间,而不影响表现(参见"Fast primer annealing")。

2x Rotor-Gene Probe RT-PCR Kit*的组分
组分 特性 优势
HotStarTaq Plus DNA聚合酶 95ºC条件下5分钟激活 在室温下建立qPCR反应体系
Rotor-Gene Probe RT-PCR缓冲液 NH4+和K+平衡组合预混液 特异性引物退火 ,确保可靠的qPCR结果
独特的Q-Bond添加剂 更快速的PCR循环,快速获得结果,一天可进行更多的反应。
Rotor-Gene RT Mix   特殊混合的反转录酶,对RNA亲和力高。 RNA可在10分钟内完成逆转录,即使是复杂的二级结构。
*包含dNTP混合液(dATP、dCTP、dGTP和dTTP)。
Procedure

即用型Rotor-Gene Probe RT-PCR Master Mix无需优化反应和扩增条件。只需在预混液中添加模板RNA、引物、探针和提供的逆转录酶混合液,然后设置PCR仪即可开始实验。试剂盒中提供的操作手册有使用说明。

水解探针(如TaqMan探针)与Rotor-Gene Probe RT-PCR Kit配合使用,在Rotor-Gene Q实时荧光定量PCR分析仪上快速、灵敏的定量,只需将引物探针混合物和模板加入预混液即可。

Applications

Rotor-Gene Probe RT-PCR Kit专用于Rotor-Gene Q实时荧光定量PCR分析仪,使用序列特异性探针采用定量一步法RT-PCR可快速定量RNA靶分子。该试剂盒也与Rotor-Gene 3000和Rotor-Gene 6000兼容。

Feature
Specifications
Applications Real-time quantification of RNA targets
Description For ultrafast quantitative real-time one-step RT-PCR using sequence-specific probes
Reaction type Real-time one-step RT-PCR
Real-time or endpoint Real-time
Sample/target type RNA
Single or multiplex Single
SYBR Green I or sequence-specific probes Sequence-specific probes
Thermal cycler Rotor-Gene Q, Rotor-Gene 3000, Rotor-Gene 6000
With or without ROX Without ROX dye

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Brochures & Guides
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Kit Handbooks
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For fast real-time PCR, two-step RT-PCR, and one-step RT-PCR using sequence-specific probes on Rotor-gene cyclers.
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