Rotor-Gene Q

For outstanding performance in real-time PCR

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Rotor-Gene Q 2plex Priority Package Plus

Cat. No. / ID: 9001862

Real-time PCR cycler with 2 channels (green, yellow), laptop computer, software, accessories: includes Priority Package with software, installation, training, 3-year warranty on parts and labor, and 3 preventive maintenance visits

InstrumentSoftware

Rotor-Gene Q

Rotor-Gene Q HRM

Rotor-Gene ScreenClust HRM Software

Multi-channel detection

2plex

5plex

6plex

Service Options

Priority Package Plus

Priority Package

System

Platform

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The Rotor-Gene Q, if used in combination with QIAGEN kits indicated for use with the Rotor-Gene Q instrument, is intended for the applications described in the respective QIAGEN kit handbooks. If the Rotor-Gene Q instrument is used with kits other than QIAGEN kits, it is the user’s responsibility to validate the performance of such product combination for any particular application. See trademarks.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

Outstanding thermal and optical performance due to rotary format
An unmatched optical range spanning UV to infrared wavelengths
State-of-the art analyses supported by user-friendly software
Low maintenance and maximum convenience due to robust design
High performance in multiple applications with QIAGEN assays

Product Details

QIAGEN's real-time PCR cycler, the Rotor-Gene Q, combines multiple optimized design features to provide the outstanding performance and reliable results that your research demands. Together with optimized QIAGEN kits for real-time PCR, the Rotor-Gene Q enables streamlined analysis for a wide range of applications. Q-Rex Software is operating and analysis software for life science qPCR applications. A choice of plug-ins lets you add new functionality without affecting established workflows.

Explore the virtual demo to learn more about the Rotor-Gene Q. Visit the Rotor-Gene Q Accessories page for Rotor-Gene Q consumables and accessories.

Performance

The Rotor-Gene Q is the only real-time cycler currently capable of deciphering the most difficult class IV SNPs by HRM. Harness the power of HRM using dedicated QIAGEN HRM Kits for applications such as genotyping (see figure "" for data from the Type-it HRM PCR Kit), quantitative methylation analysis (see figure "" for data from the EpiTect HRM PCR Kit), gene scanning, and sequence matching. The Type-it HRM PCR Kit reliably and accurately detects gene mutations and SNPs. The EpiTect HRM PCR Kit enables fast screening and accurate detection of changes in CpG methylation status of bisulfite converted DNA.

See figures

Identification of a class IV SNP.

Highly sensitive detection of methylated DNA.

Principle

Unique rotary design for outstanding performance

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:

Tube-to-tube variation ±0.02°C
Uniform detection eliminating the need for ROX reference dye
Fast ramping and negligible equilibration times for short run-times
Complete confidence in your results

Unrivaled optical range enables multiple applications

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.

Channels for optical detection

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

Expand your research with HRM

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:

A specially tuned high-intensity optical HRM channel
Thermal resolution of 0.02°C
High data-acquisition rates
Comprehensive HRM software

Minimum maintenance; maximum convenience

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:

Lifetime guarantee on highly stable LEDs, no expensive lamps or lasers to change, no gradual performance loss of light source
No optical calibration needed at installation or when the instrument is moved
No sample block to clean
No condensation or bubbles in reactions due to rotation
Small, light, and robust; simply place the instrument wherever you like

See figures

Cross-section of the Rotor-Gene Q.

Precise real-time PCR analysis.

Procedure

Flexible formats match your workflows

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.

Easy routine verification

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.

See figures

Flexible formats.

Applications

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:

Gene expression analysis
Pathogen detection
DNA methylation analysis
Genotyping and gene scanning
miRNA research

Software

Q-Rex Software

Q-Rex Software is operating and analysis software that drives Rotor-Gene Q hardware and facilitates experiment setup, data acquisition and basic and advanced analyses. The configurable nature of Q-Rex allows you to easily create the exact software configuration to meet your specific analysis needs without installing functions you will never use.

Fast and easy access to templates and recent experiments
Drag-and-drop creation of new experiments
Plug-ins to expand capacity for specific applications
Audit trail for 21CFR part 11 compliance

The software is suitable for use by the most novice researchers, while maintaining the highly complex data analysis functions required by advanced researchers.

Rotor-Gene Q Software

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.

Superior Rotor-Gene ScreenClust HRM Software available for genotyping and mutation detection using HRM analysis

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

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:

Estimation of up- and down-regulation for gene expression studies
Randomization and bootstrapping techniques
Graphical data output via whisker-box plots

Traditional relative quantification enables estimation of gene expression. However, this method does not provide statistical information that is suitable for comparing expression in groups of treated and untreated samples in a robust manner. The integrated randomization and bootstrapping methods used in REST software 2009 test the statistical significance of calculated expression ratios and can be used even when the data includes outliers.

REST software 2009 applies a mathematic model that takes into account the different PCR efficiencies of the gene of interest and reference genes. Compared to using a single reference gene, using multiple reference genes for normalization can improve the reliability of results.

Rotor-Gene Assay Manager

Rotor-Gene AssayManager is a software for routine testing. The core application is complemented by a selection of plug-ins that contain assay-type-specific analysis and visualization of the results for fully automated real-time PCR analysis. Further information is available from the following links:

Services

Have your throughput demands increased? Do you want to expand your application range or further streamline workflows? Do you need to manage more complex samples? Do you need improved connectivity?

If the answer to any of these questions is yes, then QIAGEN’s trade-in/trade-up program is just right for you!

QIAGEN makes it easy to keep up to speed with the latest automation and sample processing technologies. Simply exchange an old instrument for a new one. Improve your efficiency by trading in a competitor instrument or trading up with an older QIAGEN instrument.

Learn more about trade-in/trade-up opportunities.

Supporting data and figures

Identification of a class IV SNP.

A human A/T SNP in the AHRR7 gene was analyzed using genomic DNA from wild-type (blue), homozygous mutant (green), and heterozygous (red) samples. Experiments were performed using the Type-it HRM PCR Kit and a Rotor-Gene Q cycler with an HRM channel. Data analysis was performed with the unsupervised mode of Rotor-Gene ScreenClust HRM Software. A/T polymorphisms (class IV SNPs) are most difficult to discriminate due to minute differences between homozygote alleles (in this example, less than 0.1°C). [A] HRM raw data, [B] cluster plot. All pseudo-unknowns were correctly clustered according to genotype.

Specifications

Features	Specifications
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.)
Operating temperature	18-30ºC (64-86ºF)
Pollution level	2; Environmental class 3K2 (IEC 60721-3-3)
Humidity	10–75% (noncondensing)
Features	Dynamic range, 10 orders of magnitude
Protocols/main application on this instrument	Gene expression analysis, microRNA detection, Virus detection, SNP genotyping, SNP genotyping, High Resolution Melt analysis (HRM)
Software	Q-Rex Software, supplied on the installation CD provided
Overvoltage category	II
Heat dissipation/thermal load	Average, 0.183 kW (632 BTU/hour); Peak, 0.458 kW (1578 BTU/hour)
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
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
Place of operation	For indoor use only
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
Storage conditions	15ºC to 30ºC (59ºF to 86ºF) in manufacturer’s package; Max. 75% relative humidity (noncondensing); Environmental class 1K2 (IEC 60721-3-1)
Technology	Real-time PCR cycler
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
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
Typical run time	40 cycles in 45 min (assay dependent)
Weight	12.5 kg (27.6 lb.), standard configuration
Altitude	Up to 2000 m (6500 ft)

Service Plans

RGQ, Core Agreement

Cat. No. / ID: 9245253

One-time Rotor-Gene Q instrument repair performed at a regional repair center including shipping, labor, and spare parts for a period of one year. Instrument repair turnaround time of 7–10 business days. Includes one on-site Preventive Maintenance or Inspection Service and a 10% discount on additional repair service during the Core Agreement period.

Inquire

Resources

Fast and Integrated Screening Workflow to Assess Gene Expression of Individual Pathways

For use with the Q-Rex Software for determination of genotypes using High-Resolution Melting analysis.

For use with the Q-Rex Software to classify the genotypes of unknown samples.

For use with the Q-Rex Software to visualize fluorescence data and determine quantification cycle values based on an automatic user independent threshold algorithm.

Gene Expression Analysis Plug-in for the Q-Rex Software

For use with the Q-Rex Software v1.0 or higher to detect targets

For use with the Q-Rex Software or higher to calculate absolute concentration of targets by PCR. The plug-in is dedicated to our customers

For use with the Q-Rex Software to calculate absolute concentration of targets by PCR

For use with Q-Rex Software version 1.0 to calculate absolute concentration of targets by PCR. This user manual provides information about the functions and features of the Q-Rex Absolute Quantification HID plug-in.

Gene Expression Plug-in v3.0; for use with Q-Rex Software v2.0 for Rotor-Gene Q and QIAquant to perform relative quantitation analyses

Target detection Plug-in v2.0; for use with the Q-Rex Software v2.0 for Rotor-Gene Q and QIAquant to detect targets by PCR

For use with the Q-Rex Software v1.0 for determination of genotypes using High-Resolution Melting analysis. This user manual provides information about the functions and features of the Q-Rex HRM Analysis plug-in.

For use with the Q-Rex Software v2.0 for Rotor-Gene Q and QIAquant to classify the genotypes of unknown samples

Absolute Quantification Plug-in v3.0; for use with the Q-Rex Software v2.0 for Rotor-Gene Q and QIAquant to calculate absolute concentration of targets by PCR

For use with the Q-Rex Software v2.0 for Rotor-Gene Q and QIAquant for determination of genotypes using Melt Curve Analysis

Basic Plug-in v3.0; for use with the Q-Rex Software v2.0 for Rotor-Gene Q and QIAquant to determine quantification cycle values

For use with the Rotor-Gene Q and QIAquant instruments

Second edition — innovative tools

Now with even more applications!

Guideline for ISO 20836:2020

For use with Rotor-Gene Q instruments and Q-Rex Software version 1.0

For use with Rotor-Gene Q Software version 2.3.4

Alternative options for the optical drive (DVD-ROM drive) on the Rotor-Gene Q Notebook, Part Number 9026760

For use with Q-Rex Software to import sample information from QIAgility instruments

For use with the Rotor-Gene Q and QIAquant instruments.

Note: The Q-Rex Software is compatible only with QIAquant instruments with serial numbers that start with 3108 (produced after 11/2021).

For use on the Rotor-Gene Q. Rotor-Gene Q software 2.3.5 is compatible with Windows 7 and Windows 10 operating systems.

For use with the Q-Rex Software or higher to create new experiments quickly and easily based on existing QIAGEN kit templates.

For verification of thermal accuracy of Rotor-Gene real-time cyclers

FAQ

Please refer to section 'Guidelines for effective multiplex assays' under "Important Notes" in the Rotor-Gene Multiplex PCR Handbook for suitable combinations of reporter dyes, or visit our Multiplex real-time PCR Resource site for additional information.

FAQ ID -9028

Based on our extensive and successful testing of many QuantiTect Primer Assays with Rotor-Gene SYBR Green PCR Kits, we guarantee this.

FAQ ID -2124

Reaction volumes suitable for use on the Rotor-Gene Q are:

Rotor-Disc 100: 30 µl x 100-wells, 10-25 µl reaction volume
Rotor-Disc 72: 0.1 ml x 72-wells, 15-25 µl reaction volume
Strip Tubes 0.1 ml: 0.1 ml x 72-wells, 10-30 µl reaction volume, strips of 4 tubes and caps
PCR Tubes 0.2 ml: 0.2 ml x 36-wells, 15-50 µl reaction volume, individual tubes with caps

FAQ ID -9030

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.

FAQ ID -2087

Yes, please follow the Supplementary Protocols at the links below:

Rotor-Gene SYBR Green PCR Kit:

'Rotor-Gene software setup for the Rotor-Gene SYBR Green PCR Kit' (PCR106)

Rotor-Gene SYBR Green RT-PCR Kit:

'Rotor-Gene software setup for the Rotor-Gene SYBR Green RT-PCR Kit' (PCR107)

FAQ ID -2104

'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.

FAQ ID -2203

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.

3828

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.

FAQ ID -2204

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.

FAQ ID -2205

In the REST 2009 software, the hypothesis test performs 10,000 random reallocations of samples and controls between the groups, and counts the number of times the relative expression on the randomly assigned group is greater than the sample data.

FAQ ID -2459

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

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.

FAQ ID -2082

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.

FAQ ID -2089

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.

FAQ ID -2198

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

In the REST 2009 software, the range for the Standard Error on the results tab is 68% C.I. (confidence interval).

FAQ ID -2455

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.

FAQ ID -2079

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.

FAQ ID -2092

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.

FAQ ID -2091

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.

FAQ ID -1141

Please make data are collected in the appropriate fluorescent channel. Also check the gain is optimized.

If the issue persists, please send the original run file with extension .rex to QIAGEN Technical Service for further assistance.

FAQ ID -9023

The reaction volume to be entered in the Rotor-Gene Q software should be the sum of the PCR reaction volume and the volume of Vapor-Lock.

FAQ ID -2150

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.

FAQ ID -2083

Please send the original OTV run file to QIAGEN Technical Service for further assistance.

FAQ ID -9022

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.

FAQ ID -2122

We strongly recommend use of the dedicated Rotor-Gene Q Accessories, e.g., PCR Tubes 0.2 ml or Strip Tubes and Caps 0.1 ml provided by QIAGEN. These tubes are designed to give low background and perfectly match the Rotor-Gene Q requirements.

FAQ ID -2085

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

Yes, please choose from the Supplementary Protocols below:

QuantiFast Probe RT-PCR Kit:

QuantiFast Probe RT-PCR +ROX Vial Kit:

FAQ ID -2105

An Investigator Quantiplex Pro or Quantiplex Pro RGQ run of 40 cycles takes ca. 60 minutes.
For Investigator Quantiplex on the Rotor-Gene Q, a run of 40 cycles takes ca. 55 minutes. On the Applied Biosystems 7500 Real-Time PCR System, a run of 40 cycles takes ca. 57 minutes.

FAQ ID -2566

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.

FAQ ID -1519

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.

FAQ ID -2200

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.

FAQ ID -2202

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

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

Our RT² qPCR Primer Assays may be used on any real-time instrument. qPCR solutions are available for the most popular qPCR instrumentation, including those from QIAGEN, ABI, BioRad, Stratagene.

Instrument-specific protocols are available for selected instruments, and can be accessed at the following link: http://www.sabiosciences.com/pcrarrayprotocolfiles.php

FAQ ID -2714

The Investigator Quantiplex Kit has been validated for the Rotor-Gene Q, and the Applied Biosystems 7500 Fast and 7500 Real-Time PCR Systems.

FAQ ID -2567

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.

FAQ ID -2088

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).

FAQ ID -2201

Using the Investigator Quantiplex on the Rotor-Gene Q: All versions of the Rotor-Gene Q software can be used to run the Investigator Quantiplex.

FAQ ID -2568

No, that is not necessary. Simply use the primer concentrations specified in the protocols in the handbook supplied with your Rotor-Gene Multiplex PCR Kit.

FAQ ID -2128

For hardware related issues, please send the support package to QIAGEN Technical Service. Within the Rotor-gene Q software, click Help and select Send Support Email. In the new window, select the file that relates to the issue and email it to QIAGEN Technical Service.

FAQ ID - 9024

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

QIAGEN recommends the annual inspection service on Rotor-gene instruments, during which all application-critical modules of the Rotor-gene are inspected and tested and an OTV check is conducted. Performed tests and test results are documented in a GMP/GLP-compliant Report. In addition, the end users can perform the temperature calibration in the lab as needed using the Rotor-Disc OTV kit.

Note: The Rotor-Disc OTV kit requires the Rotor-Disc 72 Rotor and Rotor-Disc 72 locking ring.

FAQ ID -9025

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.

FAQ ID -2093

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

The Rotor-Gene Q software version v2.2 and lower versions require a computer with Windows XP or Vista operating systems. The Rotor-Gene Q software v2.3 or higher versions can run with Windows XP or Windows 7 32-bit and 64-bit operating systems.

Rotor-Gene 6000 software requires a computer with Windows XP or Vista operating system. QIAGEN will no longer support Rotor-Gene 3000 instruments and Rotor-Gene 6.0 software starting from January 1, 2014

FAQ ID -9021

Yes, simply use the assays at a final concentration of 1x with the Rotor-Gene Multiplex PCR Kit.

See trademarks.

FAQ ID -2131

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.

FAQ ID -2153

Use of endogenous control genes corrects for variation in RNA content, variation in reverse-transcription efficiency, possible RNA degradation or presence of inhibitors in the RNA sample, variation in nucleic acid recover, and differences in sample handling. The endogenous control gene ought to have consistent expression levels between samples and among treatment conditions, and ideally has a similar expression level to that of the genes of interest. Genes commonly used as references can be found at the QuantiTect Primer Assays as endogenous controls.

FAQ ID -9027

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.

FAQ ID -2123

On the Rotor-Gene Q, setting the threshold in the Yellow channel to an absolute value of 0.05 will give satisfactory results in most cases.

FAQ ID -2607

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

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.

FAQ ID -2081

The sample rotor of the Rotor-Gene Q spins continuously at a speed of 400 rpm during a run.

FAQ ID -2080

The specific features of Rotor-Gene Kits and Rotor-Gene cyclers work synergetically to enable an ultrafast-cycling protocol. We do not guarantee that the performance of the Rotor-Gene Multiplex RT-PCR Kit with the same cycling protocol will be the same on other cyclers.

FAQ ID -2142

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

Yes, all empty positions in the rotor of the Rotor-Gene Q have to be filled with empty tubes. This guarantees optimal temperature repartition in the Rotor-Gene Q chamber.

FAQ ID -9029

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.

FAQ ID -2199