Rotor-Gene Multiplex PCR Kit

For ultrafast, multiplex, real-time PCR and two-step RT-PCR on Rotor-Gene cyclers

Optimized for ultrafast, reliable results on Rotor-Gene cyclers
Sensitive detection of up to 4 targets in 1 tube
Successful multiplex PCR without the need for optimization
Reliable quantification of low- and high-abundance targets

The Rotor-Gene Multiplex PCR Kit is designed for use with the Rotor-Gene Q and other Rotor-Gene cyclers, providing ultrafast, highly reliable quantification in multiplex, real-time PCR and two-step RT-PCR using sequence-specific probes. Depending on the cycler configuration, up to 4 cDNA or gDNA targets (e.g., 1 control gene and 3 target genes) can be quantified simultaneously in the same tube. Outstanding performance is achieved through the combination of a specially optimized master mix and the unique Rotor-Gene cycler. For convenience, the master mix can be stored at 2–8°C.

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产品		货号	目录价：
Rotor-Gene Multiplex PCR Kit (80) Trial kit for 80 x 25 µl reactions: 1 ml 2x Rotor-Gene Multiplex PCR Master Mix, 2 ml RNase-Free Water		204772	询价
Rotor-Gene Multiplex PCR Kit (400) For 400 x 25 µl reactions: 3 x 1.7 ml 2x Rotor-Gene Multiplex PCR Master Mix, 2 x 2 ml RNase-Free Water		204774	询价

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Rotor-Gene Multiplex 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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Duplex, real-time two-step RT-PCR was carried out on the Rotor-Gene Q using the Rotor-Gene Multiplex PCR Kit and self-designed TaqMan assays for [A] IL8 (interleukin 8) and [B] ACTB (β-actin). Analysis of tenfold dilutions of leukocyte cDNA template from 100 ng to 1 pg provided high PCR efficiencies of around 95%. [C] The C_T values were comparable with those achieved in control singleplex reactions, demonstrating the reliability of the duplex assay.|Tenfold dilutions of human leukocyte cDNA (10 ng to 10 pg) were used as template in 4-plex, real-time PCR. Reactions were run in triplicate using either the Rotor-Gene Q and Rotor-Gene Multiplex PCR Kit or an instrument and kit from Supplier S. [A] Targets amplified, and reporter dyes of corresponding TaqMan probes. [B] C_T values obtained for all 4 targets (instrument and kit from Supplier S did not successfully detect IFNG; N.D.). Lower C_T values on the Rotor-Gene Q demonstrate detection with greater sensitivity. [C] Amplification plots for TNF using the Rotor-Gene Multiplex PCR Kit. [D] Amplification plots for TNF using a kit from Supplier S. [E] Amplification plots for IFNG using the Rotor-Gene Multiplex PCR Kit. [F] Amplification plots for IFNG using a kit from Supplier S.|[A] Q-Bond in Rotor-Gene Multiplex 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.|Rotor-Gene Multiplex PCR Kits provide a simple procedure for quantitative, multiplex, real-time PCR. In contrast to current methods, the kits eliminate the need for optimization of the concentrations of primers, Mg²⁺, and Taq DNA polymerase, even for difficult assays (e.g., assays in which the copy number of the target gene is much smaller than that for the reference gene).|[A] NH₄⁺ ions prevent nonspecific primers from annealing to the template. [B] Synthetic Factor MP, an innovative PCR additive, increases the local concentration of primers at the template. Together with K⁺ and other cations, Factor MP stabilizes specifically bound primers, allowing efficient primer extension by HotStarTaq Plus DNA Polymerase.|

Performance

With the Rotor-Gene Multiplex PCR Kit, up to 4 cDNA targets can be simultaneously and rapidly quantified in the same tube, increasing throughput and saving precious sample material (see figures "Reliable multiplex analysis" and "Reliable duplex analysis"). Genes of different expression levels are all amplified in the same tube with the same high efficiency, enabling reliable relative quantification of gene expression.

Principle

Amplifying reference and target genes in the same reaction instead of in separate reactions increases the reliability of gene quantification by minimizing handling errors. The Rotor-Gene Multiplex PCR Kit enables reliable multiplex real-time PCR quantification of cDNA targets on the Rotor-Gene Q without the need for optimization of reaction and cycling conditions (see flowchart "QIAGEN multiplex kits"). Highly specific amplification is assured through a balanced combination of K⁺ and NH₄⁺ ions, which promote specific primer annealing and enable high PCR specificity and sensitivity, while synthetic Factor MP, an innovative PCR additive specially developed for challenging multiplex PCR applications, allows different amplicons in the same reaction to all be amplified with the same high efficiency (see figure "Unique PCR buffer").

Fast cycling without compromising performance is achieved using Q-Bond, a novel PCR additive that considerably shortens cycler run times (see figure "Fast primer annealing"). In addition, the highly stringent hot-start enzyme HotStarTaq Plus DNA Polymerase is rapidly activated at the start of PCR by a brief 5-minute incubation at 95ºC.

Components of 2x Rotor-Gene Multiplex PCR Kit*
Component	Features	Benefits
HotStarTaq Plus DNA Polymerase	5 min activation at 95ºC	Set up of qPCR reactions at room temperature
Rotor-Gene Multiplex PCR Buffer	Balanced combination of NH₄⁺ and K⁺ ions	Specific primer annealing ensures reliable qPCR results
	Synthetic Factor MP	Reliable multiplexing analysis of up to 4 targets in the same tube
	Unique Q-Bond additive	Faster PCR run times enable faster results and more reactions per day

Procedure

A ready-to-use master mix eliminates the need for optimization of reaction and cycling conditions. Simply add template cDNA and primer-probe sets to the master mix and program the cycler. The handbook supplied with the kit lists recommended dyes and contains a single protocol for all multiplex qPCR assays.

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.

Applications

The Rotor-Gene Multiplex PCR Kit is optimized for fast, real-time PCR and two-step RT-PCR analysis using sequence-specific probes on the Rotor-Gene Q. It is also compatible with the Rotor-Gene 3000 and the Rotor-Gene 6000. Up to 4 cDNA or gDNA targets can be simultaneously and rapidly quantified in the same tube, increasing throughput and saving precious sample material.

For ultrafast, one-step qRT-PCR multiplex analysis of RNA targets using sequence-specific probes on Rotor-Gene cyclers, use the Rotor-Gene Multiplex RT-PCR Kit.

特点	参数
应用	Real-time quantification of genomic DNA or cDNA targets in a multiplex format
描述	For ultrafast quantitative multiplex real-time PCR and two-step RT-PCR using sequence-specific probes
反应类型	Real-time PCR and two-step RT-PCR
real-time或终点法PCR	Real-time
样本/目标类型	DNA, cDNA
单一或多重	Multiplex
SYBR Green I或序列特异性探针	Sequence-specific probes
热循环仪	Rotor-Gene Q, Rotor-Gene 3000, Rotor-Gene 6000
有/无ROX	Without ROX dye

产品介绍与指南

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	(EN) - Rotor-Gene Q - Pure Detection Now with even more applications!	显示更多

常见问题

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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	浏览
	Are Rotor-Gene Kits compatible with reaction setup using the QIAgility instrument? FAQ ID -2116	浏览
	Do the master mixes in Rotor-Gene Kits contain dUTP to allow UNG pretreatment? FAQ ID -2117	浏览
	Why is the initial activation step different for Rotor-Gene Probe, SYBR Green and Multiplex Kits? FAQ ID -2118	浏览
	What are the main differences between Rotor-Gene and QuantiTect or QuantiFast PCR Kits? FAQ ID -2119	浏览
	Will Rotor-Gene Kits also work on the Rotor-Gene 6000 and 3000 cyclers? FAQ ID -2121	浏览
	Do limiting primer concentrations need to be determined when using the Rotor-Gene Multiplex PCR Kit? FAQ ID -2128	浏览
	If housekeeping and target genes differ significantly in abundance, will they both be amplified with equal efficiency using the Rotor-Gene Multiplex PCR Kit? FAQ ID -2129	浏览
	Are probes other than TaqMan® probes, such as FRET probes, compatible with fast cycling with the Rotor-Gene Multiplex PCR Kit? FAQ ID -2130	浏览
	Are TaqMan® Gene Expression Assays from Applied Biosystems compatible with the Rotor-Gene Multiplex PCR Kit? FAQ ID -2131	浏览
	Do you have any information or guidelines regarding the choice of reference genes for real-time PCR? FAQ ID -2371	浏览
	What real-time cycler should I use for my qPCR experiments? FAQ ID -2670	浏览

试剂盒操作手册

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	(EN) - Rotor-Gene Multiplex Handbook For fast multiplex real-time PCR, two-step RT-PCR, and one-step RT-PCR using sequence-specific probes on Rotor-Gene cyclers	显示更多

技术资讯

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	(EN) - Critical factors for synthesis of cDNA for real-time PCR analysis	显示更多
	Overcoming the challenges of real-time, multiplex PCR - (EN)	显示更多

安全数据表

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

安全数据表

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	MSDS Rotor-Gene Multiplex PCR Kit (80)
	MSDS Rotor-Gene Multiplex PCR Kit (400)

图片

可靠的双重分析。

采用双重两步法real-time RT-PCR，配合Rotor-Gene Multiplex PCR Kit和自行设计的TaqMan探针在Rotor-Gene Q实时荧光定量PRC仪上检测[A] IL8（白细胞介素-8）和[B] ACTB（β-actin）。10倍稀释白细胞cDNA模板从100 ng到1 pg，PCR扩增效率高，大约为95%。[C] 获得的C_T值与那些存档的在不同反应管中反应获得的C_T值相当，由此表明该双重分析的可靠性。

可靠的多重分析。

用10倍稀释的人白细胞cDNA（10 ng到10 pg）作为模板进行4重real-time PCR。反应使用Rotor-Gene Q实时荧光定量PCR仪和Rotor-Gene Multiplex PCR Kit或供应商S提供的仪器或试剂盒，重复进行三次反应。[A]相应探针扩增的靶基因和荧光颜色。[B]4个靶基因的C_T值（供应商S提供的仪器和试剂盒不能检测IFNG；N.D.）。Rotor-Gene Q实时荧光定量PCR仪上较低的C_T值表明QIAGEN的产品有更高的灵敏度。[C]使用Rotor-Gene Multiplex PCR Kit的TNF扩增曲线。[D]使用供应商S试剂盒的TNF扩增曲线。[E]使用Rotor-Gene Multiplex PCR Kit的IFNG扩增曲线。[F]使用供应商S试剂盒的IFNG扩增曲线。

Fast primer annealing.

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

Successful Multiplex PCR-Based Genotypic Analysis without the Need for Optimization

QIAGEN multiplex kits.

Rotor-Gene Multiplex PCR Kits provide a simple procedure for quantitative, multiplex, real-time PCR. In contrast to current methods, the kits eliminate the need for optimization of the concentrations of primers, Mg²⁺, and Taq DNA polymerase, even for difficult assays (e.g., assays in which the copy number of the target gene is much smaller than that for the reference gene).

Unique Type-it Microsatellite PCR Buffer Promotes Stable and Efficient Annealing

独特的PCR缓冲液。

[A] NH₄⁺防止引物与模板的非特异性结合。[B]MP合成因子是创新的PCR添加剂，能够提高模板处局部的引物浓度，与K⁺和其他离子一起，MP合成因子能够特异性稳定结合的引物，使HotStarTaq Plus DNA Polymerase作用下的引物延伸更加高效。