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Rotor-Gene SYBR® Green RT-PCR Kit

For ultrafast, one-step qRT-PCR gene expression analysis using SYBR Green I on Rotor-Gene cyclers

  • Optimized for ultrafast, reliable results on Rotor-Gene cyclers
  • Sensitive detection of even low copy numbers
  • Accurate detection of a wide range of template amounts
  • Specially formulated, ready-to-use master mix for fast cycling
  • Guaranteed performance combined with QuantiTect Primer Assays

The Rotor-Gene SYBR Green RT-PCR Kit is designed for use with the Rotor-Gene Q and other Rotor-Gene cyclers, providing ultrafast, highly specific quantification of RNA targets with real-time one-step RT-PCR using SYBR Green I detection. 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 SYBR Green RT-PCR Kit (400)
For 400 x 25 µl reactions: 3 x 1.7 ml 2x Rotor-Gene SYBR Green RT-PCR Master Mix, 100 µl Rotor-Gene RT Mix, 2 x 2 ml RNase-Free Water
 204174
询价
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Rotor-Gene SYBR® Green RT-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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特异性检测，无需优化。|Specific primer annealing.|Fast primer annealing.|
使用10倍稀释的人白细胞RNA（100 ng到10 pg）作为模板和QuantiTect Primer Assay对BCL2（CLL/2型淋巴瘤B细胞）进行基于SYBR® Green染料的real-time one-step RT-PCR，实验重复两次。[A] Rotor-Gene Q实时荧光定量PCR分析仪和Rotor-Gene SYBR® Green RT-PCR Kit灵敏检测10 pg RNA和扩增的特异性PCR产物。[B]供应商R提供的仪器和试剂盒在Mg2+浓度优化后进行检测。但是只能检测到100 pg RNA，而且出现共扩增的非特异性产物 。|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 SYBR® Green 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.|
性能
When the Rotor-Gene SYBR Green RT-PCR Kit is used together with QuantiTect Primer Assays, highly sensitive quantification of specific PCR products is achieved without the need for optimization (see figures "Specific detection without the need for optimization").
原理

The Rotor-Gene SYBR Green RT-PCR Kit enables reliable real-time RT-PCR quantification on the Rotor-Gene Q without the need for optimization of reaction and cycling conditions. Real-time one-step RT-PCR is carried out, which means that RNA is used as template in a reaction where reverse transcription and PCR take place sequentially in the same reaction vessel. Since it is not necessary to transfer the finished RT reaction to another tube for PCR, the real-time RT-PCR procedure is streamlined, making high-throughput analysis possible.

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. Highly specific amplification is assured through a balanced combination of K+ and NH4+ ions, which promote specific primer annealing, enabling high PCR specificity and sensitivity (see figure "Specific primer annealing"). 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").

Components of 2x Rotor-Gene SYBR Green RT-PCR Kit*
ComponentFeatures Benefits
HotStarTaq Plus DNA Polymerase 5 min activation at 95ºC Set up of qPCR reactions at room temperature
Rotor-Gene SYBR Green RT-PCR Buffer Balanced combination of NH4+ and K+ ions Specific primer annealing ensures reliable qPCR results
Unique Q-Bond additive Faster PCR run times enable faster results and more reactions per day
SYBR Green I dye Yields a strong fluorescent signal upon binding double-stranded DNA Highly sensitive quantification
Rotor-Gene RT Mix   Special blend of reverse transcriptases with a high affinity for RNA RNA can be transcribed in just 10 minutes, even through complex secondary structures
* 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 RNA, primers, and the supplied reverse transcriptase mix to the master mix and program the cycler. Instructions are provided in the detailed handbook supplied with the kit.

For gene expression analysis using real-time one-step RT-PCR, the combination of the Rotor-Gene SYBR Green RT-PCR Kit with QuantiTect Primer Assays and the Rotor-Gene Q provides a complete, ready-to-run solution. 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 Rotor-Gene SYBR Green RT-PCR Kit provides rapid real-time quantification of RNA targets on the Rotor-Gene Q. The kits are also compatible with the Rotor-Gene 3000 and the Rotor-Gene 6000.

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特点
参数
应用 Real-time quantification of RNA targets
描述 For ultrafast quantitative real-time one-step RT-PCR using SYBR Green I
反应类型 Real-time one-step RT-PCR
real-time或终点法PCR Real-time
样本/目标类型 RNA
单一或多重 Single
SYBR Green I或序列特异性探针 SYBR Green I
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
(EN) - Rotor-Gene Q - Pure Detection
Now with even more applications!
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试剂盒操作手册
2
(EN) - Rotor-Gene SYBR Green Handbook
For fast real-time PCR, two-step RT-PCR, and one-step RT-PCR using SYBR Green I on Rotor-Gene cyclers
显示更多
(EN) - QuantiTect Primer Assay Handbook
For genomewide, ready-to-use real-time RT-PCR assays using SYBR Green detection
显示更多
安全数据表
1
Safety Data Sheets
Download Safety Data Sheets for QIAGEN product components.
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图片
Specific and sensitive detection using SYBR Green
特异性检测，无需优化。
使用10倍稀释的人白细胞RNA（100 ng到10 pg）作为模板和QuantiTect Primer Assay对BCL2（CLL/2型淋巴瘤B细胞）进行基于SYBR® Green染料的real-time one-step RT-PCR，实验重复两次。[A] Rotor-Gene Q实时荧光定量PCR分析仪和Rotor-Gene SYBR® Green RT-PCR Kit灵敏检测10 pg RNA和扩增的特异性PCR产物。[B]供应商R提供的仪器和试剂盒在Mg2+浓度优化后进行检测。但是只能检测到100 pg RNA，而且出现共扩增的非特异性产物 。
Specific primer annealing using a balanced combination of K+ and NH4+ ions
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 with Q-Bond
Fast primer annealing.
[A] Q-Bond in Rotor-Gene SYBR® Green 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.