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.

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

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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可靠的双重分析。|可靠的多重分析。|Fast primer annealing.|QIAGEN multiplex kits.|独特的PCR缓冲液。|
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CT值与那些存档的在不同反应管中反应获得的CT值相当,由此表明该双重分析的可靠性。|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CT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CT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。|[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+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。|
性能
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.
原理
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*
ComponentFeatures 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
* Also contains dNTP mix (dATP, dCTP, dGTP, dTTP).
操作流程

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.

应用

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

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产品介绍与指南
1
常见问题
12
试剂盒操作手册
1
For fast multiplex real-time PCR, two-step RT-PCR, and one-step RT-PCR using sequence-specific probes on Rotor-Gene cyclers
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安全数据表
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图片
Reliable duplex analysis
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CT值与那些存档的在不同反应管中反应获得的CT值相当,由此表明该双重分析的可靠性。
Reliable multiplex analysis.
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CT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CT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 with Q-Bond
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。