Rotor-Gene Q

比類ない高い感度・再現性を実現したリアルタイムPCRシステム

Products

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.

Features

遠心エアコントロール方式でサンプル間の高い温度均一性を実現（±0.02℃）
UVから赤外線領域までをカバーする広範囲にわたる光学システムに対応
使いやすいソフトウェアで最新の解析をサポート
適応性の高いデザインにより最高の簡便性と最低限のメンテナンス
QIAGENアッセイと併用し各種アプリケーションで良好な結果を実現

Product Details

QIAGEN のリアルタイムPCR 装置Rotor-Gene Qは、最高のパフォーマンスを実現するために数多くの工夫がなされており、研究ニーズにマッチした最適な結果を実現します。また、リアルタイムPCR用に至適化済みのQIAGENキットと組み合わせることにより、Rotor-Gene Q は様々なアプリケーションに効率よく対応することができます。

Performance

Rotor-Gene Q は、最も困難とされているclass IV SNP （AT 置換）のHRMによる判別が可能な唯一のリアルタイムPCR 装置です。専用のQIAGEN HRM Kit を使用したHRMはジェノタイピング（Type-it HRM PCR Kitのデータの図 "" ）、メチル化解析（EpiTect HRM PCR Kitのデータの図 ""）、遺伝子スキャニング、sequence matching などのアプリケーションにご利用いただけます。Type-it HRM PCR Kit は遺伝子変異やSNPを迅速かつ正確に検出します。EpiTect HRM PCR Kit はBisulfite変換DNAのCpG メチル化状態の変化について迅速なスクリーニングと正確な検出を実現します。

See figures

Identification of a class IV SNP.

Highly sensitive detection of methylated DNA.

Principle

遠心エアコントロール方式による卓越した性能

遠心エアコントロール方式を採用した Rotor-Gene Q は、市販されているこれまでのリアルタイムPCR装置の中で、最も精密性と機能性に富んでいます（図 " Rotor-Gene Q の断面図"）。遠心エアコントロール方式では、サンプルローターが常に遠心されていることにより、反応サンプル間の温度均一性を高く保持します（±0.02 ℃）。どのサンプルも同様に均一に検出全てのサンプルは回転しながら光源／検出器間の光路上を通過し、光源／サンプル／検出器の一定した短い光学的距離でその蛍光シグナルを迅速に得ることができます。このように温度および光学的に均一であるため、感度の高い正確かつ迅速なリアルタイムPCR解析結果が得られ（図 " 正確なリアルタイムPCR 解析"）、またサンプル間のばらつきやエッジ効果をなくします。これらは、従来のブロックベースの機器では、ブロック全体の温度勾配や複数の複雑な光経路のため避けられない問題でした。

遠心エアコントロール方式の特長:

チューブ間のばらつきは ±0.02℃

均一な検出により、ROX のようなリファレンス色素が不要

高速な加熱・冷却機構によるランニング時間の短縮

高い感度と再現性

広い光源領域で様々なアプリケーションを実現

Rotor-Gene Q は、SYBR^® Green のようなインターカレーター色素や、加水分解プローブ（TaqMan プローブ）、ハイブリダイゼーションプローブ（FRET プローブ）、Scorpion プローブ、その他幅広い蛍光波長の検出に最適です。搭載波長は、UVから赤外波長まで、最高6 チャンネルを搭載しています。さらに、ソフトウェア上での簡単な設定により、それぞれの励起光源／検出フィルターの組み合わせをカスタムで作製できるので、特殊な波長の色素にもRotor-Gene Q は対応可能です。

蛍光検出用チャンネルチャンネル励起波長 (nm)	検出波長 (nm)	検出可能な蛍光色素の例
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	Cyanine 670、Quasar 670、LightCycler	Red640、Alexa Fluor 633
Crimson	680 ± 5	712 高域	Quasar 705、LightCycler Red705、	Alexa Fluor 680
HRM	460 ± 20	510 ± 5	SYBR^® Green I、SYTO9、LC Green、	LC Green Plus+、EvaGreen

HRM 解析によるPCR 産物の識別

High Resolution Melt（HRM）解析とは、融解曲線解析を極めて精度の高い温度分解能を利用して、わずかなT_m 値の差を検出することができる技術です。HRM法は、二本鎖PCR産物を解離（融解）性に基づいて解析します。古典的な融解曲線解析に類似していますが、幅広いアプリケーション用にさらに多くの情報が得られます。PCR産物は、その塩基組成（GC 含量）、配列、長さなどに依存した産物特有の解離温度（T_m 値）を有します。Rotor-Gene Q の温度分解能は±0.02 ℃であるため、PCR 産物中に1塩基の違いがあっても、高感度に検出することができます。Rotor-Gene Q の遠心エアコントロール方式による卓越した温度均一性と光学的均一性はHRM に最適です。

Rotor-Gene Q のHRM 機能の特長：

専用のHRM 励起チャンネルを搭載

±0.02 ℃の超高温度分解能による厳密な識別

高速データ収集

自動ジェノタイピング機能を搭載したソフトウェア

最小限のメンテナンスで最大の簡便さ

Rotor-Gene Q は、日常のメンテナンスが必要なく、使いやすい設計になっています。そのため、装置本体のメンテナンスにかける時間を低減し、研究に集中することが可能です。

Rotor-Gene Q の特長:

非常に安定したLED使用のため、高価なランプやレーザーの交換が不要で、光源性能が徐々に低下することが少ない

設置時や機器移動時に必要とされる光学的補正は不要

ブロック型のような清掃が不要

回転により反応中の結露や気泡が発生しない

小さくて軽い装置なので使いやすく、ラボのどこにでも設置可能

See figures

Cross-section of the Rotor-Gene Q.

Precise real-time PCR analysis.

Procedure

スループットに応じて変更できる多様なフォーマット

Rotor-Gene Q は多様なニーズに対応するため、様々なPCR チューブフォーマットを使用できます（図 " 多様なフォーマット"）。チューブを搭載するローターを取り替えるだけでフォーマットをわずか数秒で変更できます。よりスループットの高い実験系には、Rotor-Disc が使用できます。Rotor-Disc は、円周上にサンプルウェルが配置された円形プレートです。Rotor-Disc 100を用いることで、96サンプルプレート処理と同等のスループットを実現することができ、残りの4ウェルはコントロール反応などに使用することができます。余ったウェルは、追加の反応や追加コントロールに便利に使用することができます。72ウェルのRotor-Disc 72 もあります。Rotor-Disc は、Rotor-Disc Heat Sealer を用いてプラスチックフィルムで簡単かつ迅速にシールできます。Rotor-Disc を使用する反応で必要なものは全て Rotor-Disc 100 Starter Kit あるいはRotor-Disc 72 Starter Kit に入っています。

マニュアルで反応セットアップを行なうことも、QIAGENの自動化ソリューションであるQIAgiliyを利用して迅速かつ高精度のPCRセットアップを行なうこともできます。また、QIAsymphony ASは、日常的にルーチンのPCRテストを行なうラボに最適です。どちらの装置もRotor-Gene フォーマットの自動反応セットアップを行なうことができ、サンプルリストの直接転送が可能で、リアルタイムPCRマスターミックス用の検証済みプロトコールが付属しています。

装置の定期的なバリデーションも実施可能

検査機関や診断を目的とした研究施設では、使用しているサーマルサイクラーの温度精度のバリデーションを定期的に実施することを求められる場合があります。Rotor-Gene Q では、温度精度の自動キャリブレーションを行なう専用ローター"Rotor-Disc OTV（Optical Temperature Verification）Kit"をセットしてランニングするだけで、自動的に装置の温度キャリブレーションを実施することができます。全操作は、わずか数分なので、ほかの装置のようなサービスエンジニアの出向も要請する必要もなく、費用対効果も高く効率的です。

See figures

Flexible formats.

Applications

数々のRotor-Gene Q用QIAGEN キットは、反応条件やサイクル条件の至適化の必要がなく、リアルタイムPCR アプリケーションにおいて信頼できる定量を実現します。以下の分野におけるリアルタイムPCR およびHRMに特化したキットを販売しています。

遺伝子発現解析
病原体検出
DNA メチル化解析
ジェノタイピングおよび遺伝子スキャニング
miRNA 研究

Software

定量化を可能にし、データの信頼性を向上するソフトウェア

この包括的なRotor-Gene Q software packageは、基本的から高度なアルゴリズムを搭載しており現在最高水準のリアルタイムPCR解析手順をサポートします。これにより、お客様の価値ある実験データ解析の自由度と結果に対する信頼性が増します。データはランを開始してから結果を出力するまでの全てのプロセスのステップをトラッキングできることで確実になります。

HRM解析を用いたジェノタイピングおよび変異検出用の優れたソフトウェア

Rotor-Gene ScreenClust HRM SoftwareはRotor-Gene operating softwareの拡張版です。このソフトウェアはRotor-Gene QまたはRotor-Gene 6000サイクラーから得られたHRMデータの解析のために現在使用されている最も強力なツールです。サンプルをクラスタごとに分類することで、Rotor-Gene ScreenClust HRM Softwareはジェノタイピングおよび変異スクリーニングのようなアプリケーション用のHRM解析の新しい局面を展開します。

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
Operating temperature
Pollution level
Humidity
Features
Protocols/main application on this instrument
Software
Overvoltage category
Heat dissipation/thermal load
Optical System
Power
Kits designed for this instrument
Place of operation
Samples per run (throughput)
Storage conditions
Technology
Transportation conditions
Warranty
Thermal performance
Typical run time
Weight
Altitude

Resources

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

On the Rotor-Gene Q, an Investigator Quantiplex run of 40 cycles takes approximately 48 minutes. On the Applied Biosystems 7500 Real-Time PCR System, an Investigator Quantiplex run of 40 cycles takes approximately 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