Rotor-Gene Probe PCR Kit

用于在Rotor-Gene PCR仪上使用序列特异性探针超快速进行两步法qRT-PCR

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

Rotor-Gene Probe 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两步法RT-PCR可超快速、高度灵敏的定量cDNA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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Product		Cat. no.	List price:
Rotor-Gene Probe PCR Kit (400) For 400 x 25 µl reactions: 3 x 1.7 ml 2x Rotor-Gene Probe PCR Master Mix, 2 x 2 ml RNase-Free Water		204374	528,00 €	Add to cart

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The Rotor-Gene Probe 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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Real-time PCR was carried out using two-fold dilutions of human genomic DNA (30 ng to 3.75 ng) and a self-designed TaqMan assay for IL1R2 (interleukin 1 receptor, type II). Reactions were run using either [A] the Rotor-Gene Probe PCR Kit and Rotor-Gene Q or [B] a kit and cycler from Supplier A_II (5 replicates per template dilution). The Rotor-Gene system provided lower C_T values, greater reproducibility within replicates, and greater resolution of different template dilutions.|Real-time PCR analysis of human genomic DNA was carried out using the Rotor-Gene Probe PCR Kit and Rotor-Gene Q. [A] Analysis of two-fold dilutions of DNA (from 30 ng [5000 copies] to 0.06 ng [20 copies]) using a self-designed TaqMan assay for IL1R2 (interleukin 1 receptor, type II); 5 replicates per template dilution. The average difference in C_T value between adjacent template dilutions was 1.07 cycles. [B] Analysis of 5 ng DNA using a self-designed TaqMan assay for IL1R2. From 100 replicates, the standard deviation was 0.21. [C] Analysis of 20 ng DNA using a self-designed TaqMan assay for ACTB (actin, beta). From 100 replicates, the standard deviation was 0.29.|Real-time PCR was carried out using fourfold dilutions of human genomic DNA (equivalent to 16, 666 cells down to 1 cell) and a self-designed TaqMan assay for ACTB (actin, beta). Reactions were run in triplicate using the Rotor-Gene Probe PCR Kit and Rotor-Gene Q. The Rotor-Gene system provided reliable detection over the entire range of template dilutions, with high reproducibility within each set of triplicates.|Real-time PCR was carried out using ten-fold dilutions of plasmid DNA (10⁹ to 10 copies) and a self-designed TaqMan assay for PPIA (cyclophilin A). Reactions were run in triplicate using the Rotor-Gene Probe PCR Kit and Rotor-Gene Q. The Rotor-Gene system provided reliable detection over the entire range of template dilutions as well as highly reproducible C_T values within each set of triplicates.|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 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 PCR Kit非常适合定量PCR分析（参见"Highly sensitive and reproducible detection"， "High precision in real-time PCR"），以及在大范围模板量内提供可靠的检测（参见"Reliable detection down to 1 cell"，和 "Wide dynamic range"）。

在大范围模板量内可靠检测
拷贝数	平均C_T	C_T差异
1,000,000,000	5.11	3.07
100,000,000	8.18	3.39
10,000,000	11.57	3.04
1,000,000	14.61	3.68
100,000	18.29	3.28
10,000	21.57	3.68
1,000	25.25	3.06
100	28.31	3.43
10	31.74	–
有效性	0.99
线性	1

Principle

Rotor-Gene Probe PCR Kit能够在Rotor-Gene Q实时荧光定量PCR分析仪上可靠的定量两步法RT-PCR，无需优化反应或循环条件。该试剂盒专为水解探针（如TaqMan探针）而设计。含有K⁺和NH₄⁺平衡组合的预混液促进高特异性引物退火，确保高度灵敏和特异性扩增（参见"Specific primer annealing"）。新颖的PCR添加剂Q-Bond能够在45分钟内完成循环反应，而不影响表现（参见"Fast primer annealing"）。

2x Rotor-Gene Probe 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 PCR Buffer	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
Rotor-Gene Probe PCR Buffer	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

Procedure

即用型Rotor-Gene Probe PCR Master Mix无需优化反应和循环条件。只需在预混液中添加模板cDNA、引物和探针，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 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。

为了两步法定量RT-PCR获得更加优化的结果，推荐使用QuantiTect Reverse Transcription Kit合成cDNA。该试剂盒只需20分钟即可快速合成cDNA，并可完全去除基因组DNA的污染。

Applications

Rotor-Gene Probe 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两步法RT-PCR可快速定量cDNA靶分子。该试剂盒也与Rotor-Gene 3000和Rotor-Gene 6000兼容。

在Rotor-Gene PCR仪上使用Rotor-Gene Probe RT-PCR Kit，可利用序列特异性探针对RNA靶分子进行超快速的一步法qRT-PCR基因表达分析。

Feature	Specifications
Applications	Real-time quantification of genomic DNA or cDNA targets
Description	For ultrafast quantitative real-time PCR and two-step RT-PCR using sequence-specific probes
Reaction type	Real-time PCR and two-step RT-PCR
Real-time or endpoint	Real-time
Sample/target type	DNA, cDNA
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

Brochures & Guides

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	(EN) - Rotor-Gene Q - Pure Detection Now with even more applications!	Show details

FAQs

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	How do you achieve fast cycling, yet still deliver the same performance in PCR as that achieved with standard cycling? FAQ ID -1430	View
	Are Rotor-Gene Kits compatible with reaction setup using the QIAgility instrument? FAQ ID -2116	View
	Do the master mixes in Rotor-Gene Kits contain dUTP to allow UNG pretreatment? FAQ ID -2117	View
	Why is the initial activation step different for Rotor-Gene Probe, SYBR Green and Multiplex Kits? FAQ ID -2118	View
	What are the main differences between Rotor-Gene and QuantiTect or QuantiFast PCR Kits? FAQ ID -2119	View
	Will Rotor-Gene Kits also work on the Rotor-Gene 6000 and 3000 cyclers? FAQ ID -2121	View
	How do Rotor-Gene Probe Kits compare with QuantiFast Probe Kits? FAQ ID -2125	View
	Are TaqMan® Gene Expression Assays from Applied Biosystems compatible with Rotor-Gene Probe Kits? FAQ ID -2126	View
	Do you have any information or guidelines regarding the choice of reference genes for real-time PCR? FAQ ID -2371	View
	What real-time cycler should I use for my qPCR experiments? FAQ ID -2670	View
	What is the threshold cycle or Ct value? FAQ ID -2682	View

Kit Handbooks

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	(EN) - Rotor-gene Probe Handbook For fast real-time PCR, two-step RT-PCR, and one-step RT-PCR using sequence-specific probes on Rotor-gene cyclers.	Show details

Technical Information

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	(EN) - Critical factors for synthesis of cDNA for real-time PCR analysis	Show details

Safety Data Sheets

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	Safety Data Sheets Download Safety Data Sheets for QIAGEN product components.	View

Safety Data Sheets

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	MSDS Rotor-Gene Probe PCR Kit (400)

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Highly sensitive and reproducible detection.

High precision in real-time PCR.

Real-time PCR analysis of human genomic DNA was carried out using the Rotor-Gene Probe PCR Kit and Rotor-Gene Q. [A] Analysis of two-fold dilutions of DNA (from 30 ng [5000 copies] to 0.06 ng [20 copies]) using a self-designed TaqMan assay for IL1R2 (interleukin 1 receptor, type II); 5 replicates per template dilution. The average difference in C_T value between adjacent template dilutions was 1.07 cycles. [B] Analysis of 5 ng DNA using a self-designed TaqMan assay for IL1R2. From 100 replicates, the standard deviation was 0.21. [C] Analysis of 20 ng DNA using a self-designed TaqMan assay for ACTB (actin, beta). From 100 replicates, the standard deviation was 0.29.

Reliable detection down to 1 cell.

Real-time PCR was carried out using fourfold dilutions of human genomic DNA (equivalent to 16, 666 cells down to 1 cell) and a self-designed TaqMan assay for ACTB (actin, beta). Reactions were run in triplicate using the Rotor-Gene Probe PCR Kit and Rotor-Gene Q. The Rotor-Gene system provided reliable detection over the entire range of template dilutions, with high reproducibility within each set of triplicates.

Wide dynamic range.

Real-time PCR was carried out using ten-fold dilutions of plasmid DNA (10⁹ to 10 copies) and a self-designed TaqMan assay for PPIA (cyclophilin A). Reactions were run in triplicate using the Rotor-Gene Probe PCR Kit and Rotor-Gene Q. The Rotor-Gene system provided reliable detection over the entire range of template dilutions as well as highly reproducible C_T values within each set of triplicates.

Specific primer annealing.

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.

Fast primer annealing.

[A] Q-Bond in Rotor-Gene Probe 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.