Rotor-Gene Probe PCR Kit

For ultrafast, real-time PCR and two-step RT-PCR using sequence-specific probes

Features

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

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Rotor-Gene Probe PCR Kit (400)

Cat. No. / ID: 204374

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
$489.00
This product will be discontinued as of June 30, 2022 or until stocks last. We recommend 208254.
Add to cart
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.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Product Details

The Rotor-Gene Probe PCR Kit is designed for use with the Rotor-Gene Q and other Rotor-Gene cyclers, providing ultrafast, highly sensitive quantification of gDNA and cDNA targets with real-time PCR and two-step RT-PCR using sequence-specific probes. 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.

 

IMPORTANT NOTE: This product is phasing-out until June 30, 2021, and afterward, available only until stocks last. Visit the product page of the successor kit to view improved features or to request a trial kit.

 

For more information and FAQs on this transition, visit: www.qiagen.com/PCRresource.

Performance

The Rotor-Gene Probe PCR Kit is well suited for use in quantitative PCR analysis (see figures " Highly sensitive and reproducible detection", " High precision in real-time PCR"), and provides reliable detection over a wide range of templates (see table and figures " High precision in real-time PCR", " Reliable detection down to 1 cell", and " Wide dynamic range").

 

Reliable detection over a wide dynamic range
CopiesMean CTCT difference
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
Efficiency 0.99
Linearity 1
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).

See figures

Principle

The Rotor-Gene Probe PCR Kit enables reliable real-time two-step RT-PCR quantification on the Rotor-Gene Q without the need for optimization of reaction and cycling conditions. The kit is designed for use with hydrolysis probes (e.g., TaqMan® 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 enables cycler run times of as low as 45 minutes (see figure " Fast primer annealing").

Components of 2x Rotor-Gene Probe PCR Kit*
Component Features Benefits
HotStarTaq Plus DNA Polymerase 5 min activation at 95ºC Set up of qPCR reactions at room temperature
Rotor-Gene Probe 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
* Also contains dNTP mix (dATP, dCTP, dGTP, dTTP).
See figures

Procedure

The ready-to-use Rotor-Gene Probe PCR Master Mix eliminates the need for optimization of reaction and cycling conditions. Just add template DNA, primers, and probe to the master mix and program the cycler. Instructions are provided in the detailed handbook supplied with the kit.

Hydrolysis probes (e.g., TaqMan® probes) can be used in combination with the Rotor-Gene Probe PCR Kit on the Rotor-Gene Q for fast and sensitive quantification — simply add the primer-probe mix and template to the master mix.

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 Probe PCR Kit is for fast, real-time PCR and two-step RT-PCR of gDNA or cDNA targets using sequence-specific probes on the Rotor-Gene Q. It is also compatible with Rotor-Gene 3000 and the Rotor-Gene 6000 cyclers.

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

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsReal-time quantification of genomic DNA or cDNA targets
DescriptionFor ultrafast quantitative real-time PCR and two-step RT-PCR using sequence-specific probes
Real-time or endpointReal-time
Reaction typeReal-time PCR and two-step RT-PCR
Sample/target typeDNA, cDNA
SYBR Green I or sequence-specific probesSequence-specific probes
Single or multiplexSingle
Thermal cyclerRotor-Gene Q, Rotor-Gene 3000, Rotor-Gene 6000
With or without ROXWithout ROX dye

Resources

Kit Handbooks (1)
For fast real-time PCR, two-step RT-PCR, and one-step RT-PCR using sequence-specific probes on Rotor-gene cyclers.
Safety Data Sheets (1)
Download Safety Data Sheets for QIAGEN product components.
Brochures & Guides (1)
Now with even more applications!

FAQ

Do you have any information or guidelines regarding the choice of reference genes for real-time PCR?

Yes, please visit our website section 'Using endogenous control genes in real-time RT-PCR' for general information. It provides a list of relative gene expression levels for commonly used human and mouse reference genes.

We offer a set of ready-to-order control genes for use in SYBR Green based as well as probe based real-time RT-PCR.

In addition, you may want to refer to the following citations on reference gene selection for quantitative real-time PCR:

• Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, DePaepe A, Speleman F [2002]: Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 2002, 3:0034.

• Radonic A, Thulke S, Mackay IM, Landt O, Siegert W, Nitsche A., 2004. Guideline to reference gene selection for quantitative real-time PCR. Biochem Biophys Res Commun. 313(4): 856-62.

• Katrien Smits,Karen Goossens, Ann Van Soom, Jan Govaere, Maarten Hoogewijs, Emilie Vanhaesebrouck,Cesare Galli, Silvia Colleoni, Jo Vandesompele, and Luc Peelman [2009]Selection of reference genes for quantitative real-time PCR in equine in vivo and fresh and frozen-thawed in vitro blastocysts. BMC Res Notes. Dec 11;2:246.

FAQ ID -2371
How do Rotor-Gene Probe Kits compare with QuantiFast Probe Kits?

Rotor-Gene Probe Kits are specially developed for Rotor-Gene cyclers. The unique rotary system of the cyclers combined with the kits’ proprietary buffer system enable ultrafast cycling. Rotor-Gene Probe Kits do not contain ROX dye.

 

FAQ ID -2125
Do the master mixes in Rotor-Gene Kits contain dUTP to allow UNG pretreatment?

No. The master mixes in Rotor-Gene Kits contain dTTP instead of dUTP. If UNG treatment is required, we recommend using QuantiTect +UNG Kits. QuantiTect Kits are also compatible with the Rotor-Gene Q; however, the kits require a significantly longer cycling time.

 

 

FAQ ID -2117
Why is the initial activation step different for Rotor-Gene Probe, SYBR Green and Multiplex Kits?

The buffer composition, which affects the initial reactivation of HotStarTaq Plus DNA Polymerase, has been optimized for each respective Rotor-Gene Kit.

 

FAQ ID -2118
What is the threshold cycle or Ct value?
The Ct or threshold cycle value is the cycle number at which the fluorescence generated within a reaction crosses the fluorescence threshold, a fluorescent signal significantly above the background fluorescence. At the threshold cycle, a detectable amount of amplicon product has been generated during the early exponential phase of the reaction. The threshold cycle is inversely proportional to the original relative expression level of the gene of interest.
FAQ ID -2682
Are TaqMan® Gene Expression Assays from Applied Biosystems compatible with Rotor-Gene Probe Kits?

Yes, use the assays at a final concentration of 1x with Rotor-Gene Probe Kits on the Rotor-Gene Q cycler.

See trademarks.  

FAQ ID -2126
How do you achieve fast cycling, yet still deliver the same performance in PCR as that achieved with standard cycling?

Our unique multiplex PCR buffer system with ammonium and potassium ions and Factor MP has been further optimized in QuantiFast and Rotor-Gene Kits. We have also discovered Q-Bond, a buffer component which supports the rapid formation of the polymerase–primer–template complex, leading to reduced annealing times.

FAQ ID -1430
What real-time cycler should I use for my qPCR experiments?
There are several manufacturers of high-quality real-time cyclers. These include QIAGEN's Rotor-Gene Q, Applied Biosystems, BioRad, Stratagene, Eppendorf, Roche, TaKaRa, Fluidigm, and Cepheid. The important thing to keep in mind is that, once you select an instrument to use, you must use compatible Rotor-Gene Discs and tubes, 96 or 384 well plates, and qPCR master mixes that are optimized for use in that particular instrument.  For example, QIAGEN's Rotor-Gene SYBR Green, Probe, and Multiplex real-time master mixes.  
FAQ ID -2670
Are Rotor-Gene Kits compatible with reaction setup using the QIAgility instrument?

The majority of the Rotor-Gene Kit data shown in our literature has been generated with the help of the QIAgility instrument. We did not observe any problems during the pipetting steps.

 

FAQ ID -2116
What are the main differences between Rotor-Gene and QuantiTect or QuantiFast PCR Kits?

Rotor-Gene Kits are specifically developed for the Rotor-Gene Q PCR Cycler. The unique rotary system of the cycler combined with the kits’ proprietary buffer system enable ultrafast cycling. Rotor-Gene Kits do not contain ROX dye since no normalization to a passive reference is required. Also, Rotor-Gene Kits do not contain dUTP; therefore, UNG pretreatment is not possible.

 

FAQ ID -2119
Will Rotor-Gene Kits also work on the Rotor-Gene 6000 and 3000 cyclers?

Yes. Rotor-Gene Kits will also work on the Rotor-Gene 6000 and Rotor-Gene 3000 PCR cyclers with the cycling conditions specified in the Rotor-Gene kit handbooks.

 

FAQ ID -2121