Cat. No. / ID: 9001861
The unique centrifugal rotary design of the Rotor-Gene Q makes it the most precise and versatile real-time PCR cycler currently available (see figure " Cross-section of the Rotor-Gene Q"). Each tube spins in a chamber of moving air, keeping all samples at precisely the same temperature during rapid thermal cycling. Detection is similarly uniform. When each tube aligns with the detection optics, the sample is illuminated and the fluorescent signal is rapidly collected from a single, short optical pathway. This thermal and optical uniformity results in sensitive, precise, and fast real-time PCR analysis (see figure " Precise real-time PCR analysis"). It also eliminates sample-to-sample variations and edge effects. These are unavoidable in traditional block-based instruments due to temperature gradients across the block and multiple, complex optical pathways.
The rotary design delivers:
Whether your assay is based on intercalating dyes such as SYBR® Green, probes such as hydrolysis (TaqMan), hybridization (FRET), Scorpion probes, or other multiplex chemistries, the Rotor-Gene Q meets your requirements. With up to 6 channels spanning UV to infrared wavelengths, the cycler delivers the widest optical range currently available (see table). In addition, the software allows you to create new excitation/detection wavelength combinations, which means that the Rotor-Gene Q is compatible with dyes you may use in the future.
|Channel||Excitation (nm)||Detection (nm)||Examples of fluorophores detected|
|Blue||365 ± 20||460 ± 20||Marina Blue, Edans, Bothell Blue, |
Alexa Fluor 350, AMCA-X
|Green||470 ± 10||510 ± 5||FAM, SYBR® Green I, Fluorescein, |
EvaGreen, Alexa Fluor 488
|Yellow||530 ± 5||557 ± 5||JOE, VIC, HEX, TET, MAX, CAL Fluor Gold 540, Yakima Yellow|
|Orange||585 ± 5||610 ± 5||ROX, CAL Fluor Red 610, Cy 3.5, |
Texas Red, Alexa Fluor 568
|Red||625 ± 5||660 ± 10||Cy5, Quasar 670, LightCycler |
Red640, Alexa Fluor 633
|Crimson||680 ± 5||712 high pass||Quasar 705, LightCycler Red705, |
Alexa Fluor 680
|HRM||460 ± 20||510 ± 5||SYBR® Green I, SYTO9, LC Green, |
LC Green Plus+, EvaGreen
High-resolution melting analysis (HRM) is a closed-tube, post-PCR analysis that has raised enormous scientific interest. HRM characterizes double-stranded PCR products based on their dissociation (melting) behavior. It is similar to classical melting curve analysis, but provides far more information for a wider range of applications. PCR products can be discriminated according to sequence, length, GC content, or strand complementarity, down to single base-pair changes. Previously unknown and even complex sequence variations can be readily detected and characterized in a robust and straightforward way. The rotary design of the Rotor-Gene Q and its outstanding thermal and optical performance are highly suited to HRM.
The HRM option for the Rotor-Gene Q includes:
The Rotor-Gene Q is engineered to reduce the need for maintenance and to maximize ease of use. This saves time and costs, and allows you to focus on your research; not on keeping the cycler up and running.
Convenient features of the Rotor-Gene Q include:
The Rotor-Gene Q supports multiple PCR tube formats to suit a range of needs (see figure " Flexible formats"). Changing the format, by simply switching the snap-fit metal rotor that holds the tubes, takes just seconds. As well as tubes, Rotor-Discs are available, which offer accelerated setup and higher throughput. Rotor-Discs are circular plates of vertically oriented reaction wells. The Rotor-Disc 100 is the equivalent of a 96-well plate with an additional 4 reference wells. These extra wells can be conveniently used for more reactions or additional controls. Alternatively, the Rotor-Disc 72 has 72 wells. Rotor-Discs can be quickly and easily sealed with plastic film using a Rotor-Disc Heat Sealer. For all you need to run reactions using Rotor-Discs, choose the Rotor-Disc 100 Starter Kit or the Rotor-Disc 72 Starter Kit.
You can perform manual reaction setup, or take advantage of QIAGEN's automated solutions for reaction setup. The QIAgility is cost-effective and delivers rapid, high-precision PCR setup, while the QIAsymphony AS is ideal for laboratories performing routine PCR tests on a day-to-day basis. Both instruments perform automated reaction setup in Rotor-Gene formats, allow direct transfer of sample lists, and are supplied with verified protocols for real-time PCR master mixes.
Laboratories may often want to verify thermal accuracy. For most cyclers, this requires interaction with a service engineer. With the Rotor-Gene Q, this is not necessary. Instead, the easy-to-use, cost-effective Rotor-Disc OTV (Optical Temperature Verification) Kit automates accuracy testing. The full procedure takes only a couple of minutes.
A range of QIAGEN kits for the Rotor-Gene Q enables reliable quantification in all your real-time PCR applications without the need for optimization of reaction and cycling conditions. Kits for real-time PCR and HRM applications are available for:
Q-Rex Software is a new operating and analysis software for the Rotor-Gene Q, providing several unique new features that promote a more user-friendly interface to help streamline your qPCR workflow. The software is suitable for use by the most novice researchers, while maintaining the highly complex data analysis functions required by advanced researchers.
The comprehensive Rotor-Gene Q software package supports all current state-of-the art real-time analysis procedures from basic to advanced algorithms. This provides complete freedom to analyze your valuable experimental data and increases the reliability of your results. Data security is assured and all process steps are trackable from starting the run to exporting the results.
Rotor-Gene ScreenClust HRM Software is an extension to the Rotor-Gene operating software. This software is the most powerful tool currently available for analysis of HRM data from the Rotor-Gene Q or Rotor-Gene 6000 cycler. By grouping samples into clusters, Rotor-Gene ScreenClust HRM Software opens a new dimension in HRM analysis for applications such as genotyping and mutation screening.
REST software 2009 is a standalone software tool for analysis of gene expression data from quantitative real-time PCR experiments. REST software 2009 is available for download under the "Resources" tab, and provides valuable analysis, including:
|Protocols/main application on this instrument||Gene expression analysis, microRNA detection, Virus detection, SNP genotyping, SNP genotyping, High Resolution Melt analysis (HRM)|
|Heat dissipation/thermal load||Average, 0.183 kW (632 BTU/hour); Peak, 0.458 kW (1578 BTU/hour)|
|Weight||12.5 kg (27.6 lb.), standard configuration|
|Features||Dynamic range, 10 orders of magnitude|
|Transportation conditions||–25ºC to 60ºC (–13ºF to 140ºF) in manufacturer’s package; Max. 75% relative humidity (noncondensing); Environmental class 2K2 (IEC 60721-3-2)|
|Warranty||1 year on instrument; lifetime guarantee on excitation LEDs|
|Operating temperature||18-30ºC (64-86ºF)|
|Place of operation||For indoor use only|
|Storage conditions||15ºC to 30ºC (59ºF to 86ºF) in manufacturer’s package; Max. 75% relative humidity (noncondensing); Environmental class 1K2 (IEC 60721-3-1)|
|Samples per run (throughput)||Tubes 0.2 ml; Strip Tubes 0.1 ml (4 tubes); Rotor-Disc 72; Rotor-Disc 100; Up to 100 samples per run using a Rotor-Disc 100|
|Typical run time||40 cycles in 45 min with the QIAGEN RG Kits (assay dependent)|
|Dimensions||Width, 37 cm (14.6 in.); Height, 28.6 cm (11.3 in.); Depth (without cables), 42 cm (16.5 in.); Depth (door open), 53.8 cm (21.2 in.)|
|Power||100–240 V AC, 50–60 Hz, <520 VA (peak); Power consumption <60 VA (standby); Mains supply voltage fluctuations are not to exceed 10% of the nominal supply voltages; F5a 250 V fuse|
|Kits designed for this instrument||artus QS-RGQ Kits (not available in all countries), RG SYBR Green PCR Kits; RG SYBR Green RT-PCR Kit; RG Probe PCR Kits; RG Probe RT-PCR Kit; RG Multiplex PCR Kit|
|Thermal performance||Temperature range, 35 to 99ºC; Temperature accuracy, ±0.5ºC (type, measured 30 seconds after clock start); Temperature resolution, ±0.02ºC (smallest programmable increment); Temperature uniformity, ±0.02ºC; Ramp rate (peak ramp rates, air), >15ºC/s heating, >20ºC/s cooling|
|Pollution level||2; Environmental class 3K2 (IEC 60721-3-3)|
|Software||Rotor-Gene Q software, supplied on the installation CD provided|
|Optical System||Up to 6 channels spanning UV to infra-red wavelengths; Excitation sources: High energy light-emitting diodes; Detector: Photomultiplier; Acquisition time: 4 s. The software allows to create new excitation/detection wavelength combinations|
|Technology||Real-time PCR cycler|
|Altitude||Up to 2000 m (6500 ft)|
The Rotor-Gene Q software allows creation of new excitation/detection wavelength combinations, which means that Rotor-Gene Q will work with dyes you may want to use in the future.
Yes, please follow the Supplementary Protocols at the links below:
'Probability' is the likelihood or chance that a sample is a member of each available cluster. The combined probabilities of a single sample add up to 1.00.
'Typicality' in the Rotor-Gene ScreenClust HRM analysis is a measure of how well a sample fits into its assigned cluster. It can also be seen as a measure of how far away a sample is from the centre of the cluster. Typicality values range from 0 to 1, the higher the value the closer it is to the centre of its assigned cluster. If a sample has a typicality value of 0.5, it means that approximately half of all samples within that cluster will be closer to the centre and the other half will be further away. In reality, this might not be the case as small samples numbers can have skewed distributions.
RNA purified with the QIAamp Viral RNA Mini Kit has been used for quantification by qPCR with QuantiTect Probe RT-PCR Kit on QIAGEN Rotor-Gene Q.
Clusters in Rotor-Gene ScreenClust HRM Software are graphically represented by ellipses/ellipsoids which act as a visual aid. They are not designed to cover all of the samples. They are a good tool for compare differences between clusters. To judge how well individual samples fit within their clusters use the typicality scores.
The controls define the centre of the clusters in supervised mode using Rotor-Gene ScreenClust HRM Software. If the control samples lie on the fringe of the cluster then the cluster centre can be shifted away from the bulk of the samples within the cluster. Controls should provide a good representation of the expected behavior of unknown samples. If this is not the case the experimental setup should be re-evaluated.
Both types of tubes run on the Rotor-Gene Q, and can be labeled on the top without any interference with data acquisition, because signal is detected from the bottom of the tube. Labeling is helpful and can prevent possible sample mix up.
There are several features available to allow exporting data across platforms to a variety of software packages. Sample names can easily be imported into the Rotor-Gene Q Excel sample sheets. By using a function "Export to Excel" the software automatically exports any results table in the software to Excel. Furthermore, reports can now be exported as Word files, which make the data compatible with any Microsoft office/Mac application. Reports can also be saved or sent in HTML format. All figures and graphs can be exported as Jpeg or Bitmap files for use in any desktop publishing software.
Normalization using Rotor-Gene ScreenClust HRM Software is required since HRM melt curves can have different starting points. Therefore the scale of each melt curve can be different. Comparison can only occur if all samples are on the same scale, so each curve needs to be normalized.
No. The fixed optical path ensures uniform illumination and detection from sample to sample eliminating the need to use a reference dye such as ROX on the Rotor-Gene Q.
Yes. The sample sheet can be amended during or after a PCR run by activating the 'Edit samples' button in the Rotor-Gene Q software. The sample sheet will open and may be modified.
Yes, data can be analyzed on the Rotor-Gene Q while a run is in progress, allowing to save time for planning and setting up new experiments. Multiple copies of the Rotor-Gene Q software can be opened simultaneously during a run allowing analysis of previous experiments while the system is running.
QuantiTect Primer Assays are primer pairs designed and bioinformatically validated specifically for real-time RT-PCR with SYBR Green detection. To find a primer assay for your target gene of interest, please visit our GeneGlobe data base.
For best results, we strongly recommend using QuantiTect Primer Assays in combination with QIAGEN's products for SYBR Green-based Real-Time PCR and RT-PCR.
At the selected data acquisition step of the thermal cycle on the Rotor-Gene Q, the fluorescent signal intensity is measured 20 times for each sample as they spin past the detector. Tubes on a rotor spin past the excitation/detection optics every 150 milliseconds. The average of the 20 readings is then taken as the fluorescence of the sample at that cycle number.
This type of cycling allows a significant reduction in cycling time for Rotor-Gene SYBR Green PCR Kits. It is more effective than reducing the individual times for annealing and extension.
The buffer composition, which affects the initial reactivation of HotStarTaq Plus DNA Polymerase, has been optimized for each respective Rotor-Gene Kit.
Yes, please choose from the Supplementary Protocols below:
The Rotor-Gene Q instrument can be used with two different rotor formats: using tubes or rotor-dics. Tubes can be run in 36 and 72-well rotors and rotor-discs in Rotor-Disc 72 and Rotor-Disc 100 rotors. When programming the temperature profile please make sure the correct rotor type is selected.
Principal component analysis is a well-established method of data analysis for multivariate data sets, such as obtained from, e.g., microarray analysis or image analysis. However, it is new in ScreenClust for HRM data. Principal Components (PCs) are extracted from the residuals plot so that the first Principle Component (PC1) represents the greatest variability or difference between all samples. The second (PC2) represents the regions of difference not already present in PC1. The third (PC3) represents differences not in PC1 and PC2.
The supervised mode in the Rotor-Gene ScreenClust HRM Software is designed for data sets with a known number of clusters where each cluster has defined controls. All samples will be grouped into one of the defined clusters.
The unsupervised mode is used if there are no controls for each cluster or if the number of clusters is not known. Based on the data presented, ScreenClust will return the recommended number of clusters and whether to separate the data in 2D or 3D. A user may choose to change either of both of these values.
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.
Instrument-specific protocols are available for selected instruments, and can be accessed at the following link: http://www.sabiosciences.com/pcrarrayprotocolfiles.php
Based on the defined standards present, the automatic threshold function of the Rotor-Gene Q scans through all the possible threshold levels until the best fit is determined. This is defined as the R value or correlation coefficient, which is maximized to most closely approach 1.0. If there is no standard present, the threshold can also be set manually.
Clusters analyzed in the Rotor-Gene ScreenClust HRM Software are groups of samples with the same melt characteristics. For example, in a single nucleotide polymorphism (SNP) analysis the clusters may represent the genotypes 'wild type', 'mutant' and 'heterozygote'. ScreenClust is designed to group samples into clusters after they are separated based on differences in their melt curves. The clusters can either be defined by control samples (supervised mode) or ScreenClust can determine the appropriate number of clusters (unsupervised mode).
Yes. If the reaction efficiency between two PCR runs is not expected to vary, importing a standard curve from a previous run allows to estimate concentrations when a standard curve for the current run is not available. Curves can be imported from another channel, or from another run by clicking on 'Import Curve' in the Rotor-Gene Q software. Standard curves can be adjusted such that only the efficiency of the source curve is imported into the current run. Whether a standard curve should be adjusted depends on the PCR chemistry used. To adjust a standard curve, use a reference with a known concentration in the target run.
Vapor-Lock is fully compatible with the QIAgility instrument for high-precision, automated reaction setup. It is also highly suited for use with the Rotor-Gene Q cycler. For support to program your QIAgility instrument for use with Vapor-Lock, please contact your local QIAGEN Technical Support Department.
We have performed numerous tests comparing the performance of Rotor-Gene SYBR Green PCR Kits and QuantiTect Kits with QuantiTect Primer Assays. Due to the optimized ion concentrations in the PCR buffers, both perform equally well with QuantiTect Primer Assays and do not require any adjustment of primer concentration.
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.
No. The speed at which the samples spin on the Rotor-Gene Q is not high enough to apply any significant centrifugal force on them.
The sample rotor of the Rotor-Gene Q spins continuously at a speed of 400 rpm during a run.
Once all melt curves are normalized, they are on a comparable basis. To make the information within each curve more useful for comparison, each curve is differentiated. The Residuals Plot is a plot of the difference between each sample and the composite median of all the samples after differentiation. The Residuals Plot of the Rotor-Gene ScreenClust HRM Software is different from a "Difference Plot" known from standard HRM software packages.