QIAcuityデジタルPCRシステム

ナノプレートを使用するデジタルPCRアプリケーション向け

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QIAcuity One, 2plex Device

Cat. No. / ID: 911001

最大2色の蛍光色素を検出する1プレートタイプのデジタルPCR装置、ローラー、USBフラッシュメモリー、およびQIAcuity Software Suite。予防保守点検訪問1回分を含みます。人件費、交通費、部品費について1年間保証。

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Format

One

Four

Eight

Multi-channel detection

2plex

5plex

Service Options

Device

System

System FUL-2

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QIAcuityは分子生物学的アプリケーションでの使用を想定しています。本製品を疾病の診断、予防、治療に使用することはできません。そのため臨床利用（診断、予後、治療、または血液バンク）のための製品の性能特性は不明です。

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Features

完全統合型のシステム
スケーラブルなフォーマット（装置プレート1枚、4枚、8枚）
一歩進んだマルチプレックス性能（最大5プレックス）
柔軟性に富んだサンプルスループット
わずか2時間ですべての結果を提供

Product Details

QIAcuityデジタルPCRシステムは、突然変異検出、コピー数多型（Copy Number Variation, CNV）、遺伝子発現研究、遺伝子編集分析、その他多くの用途で正確なマルチプレックス定量を実現するように設計されています。ナノプレートを使用する本システムは、サンプル分配、熱サイクル、およびイメージングの標準的なdPCRワークフローを最小限の手作業時間で実行する、シームレスに統合された完全自動化プラットフォームです。

本システムはQIAcuity Nanoplatesおよびアクセサリーと組み合わせて使用します。

QIAcuityについての詳しい情報は、バーチャルデモをご覧ください。

Performance

QIAcuityデジタルPCRシステムは、すべてのラボへ絶対定量をお手頃な価格で提供します。サンプル分配、熱サイクル、イメージングの全デジタルPCRワークフローを1つに統合、簡略化して最小限のハンズオン時間で実行する自動化装置です。また、現行のqPCRアッセイをQIAcuityデジタルPCRシステムに適用することも簡単です。qPCRの場合、プレート操作を変える必要はなく、迅速なアッセイセットアップと約2時間での迅速な結果の提供をお約束します。

QIAcuity装置 – 機能と仕様

機能	QIAcuity One	QIAcuity Four	QIAcuity Eight
プレート処理数	1	4	8
検出チャンネル（マルチプレックス）	2または5	5	5
サーモサイクラー	1	1	2
結果までの時間	約2時間	最初のプレートは約2時間その後のプレートは1枚あたり1時間以内	最初のプレートは2時間以内その後のプレートは1枚あたり30分以内
スループット（1日のサンプル処理数）	最大384（96ウェル）最大96（24ウェル）	最大672（96ウェル）最大168（24ウェル）	最大1248（96ウェル）最大312（24ウェル）

Principle

分注およびロード、実験、結果解析というわずか3つのステップで、約2時間でdPCRの結果が得られます。

ナノプレートでのdPCRの原理はこちらで説明しています。

Procedure

qPCR実験と同様に、サンプル調製では、マスターミックス、プローブ、およびプライマーを96または24ウェルナノプレートへ加え、その後サンプルを加えます。このシステムは、サンプル分配、熱サイクル、イメージングを1つに統合した完全自動化装置です。2時間未満でサンプルの結果が得られます。Software Suiteは、ターゲットシーケンスのマイクロリットルあたりのコピー数で濃度を測定するだけでなく、陽性サンプルやNTCなどの品質コントロールも解析できます。この解析は、同じローカルエリアネットワーク（LAN）内の離れた場所のコンピューターでも行うことができます。

Applications

QIAcuity装置は、QIAcuity NanoplateｓおよびQIAcuity PCR Kitと組み合わせて、次のデジタルPCRアプリケーションに利用できます。

希少突然変異の検出
コピー数多型解析
遺伝子発現解析
病原体検出
遺伝子型決定
miRNA研究

Software

QIAcuity Software Suiteは装置に同梱されており、個々のコンピューターにインストールします。コンピューターを装置に直接接続、または既存のローカルエリアネットワーク（LAN）を使用して、1台または複数のQIAcuity装置を制御できます。デジタルPCR実験において、QIAcuity Software Suiteでサンプルや反応液を定義してナノプレートに割り当て、QIAcuity装置に情報を転送できます。ラン終了後は、データ解析とレポート作成が可能です。解析のためデータを外部へエクスポートすることができます。本ソフトウェアには、繰り返し使えるプレートレイアウトや簡単にアクセスできるプレートランパラメーターの設定などのテンプレート機能があり、デジタルPCR実験をさらに拡張させることができます。

ローカルエリアネットワークに統合すれば、そのコンピューターはサーバーとしてQIAcuity Software Suiteの機能をホストし、クライアントとして機能する他のコンピューターにLANを経由してアクセスできるようになります。これにより、複数のユーザーが他の部屋やオフィスからソフトウェアにアクセスしたり、複数のコンピューターにソフトウェアをインストールしなくても標準的なブラウザを介してデータを解析したり、インターネット接続を経由してデータのやり取りができるようになります。

Services

QIAGENの多様なサービスソリューションなら機器を安心してご利用いただけます。ニーズに合ったサービス契約の詳細をご覧ください。

Supporting data and figures

プレート方式のシンプルで迅速なワークフロー

パーティショニング、熱サイクル、イメージングを1台で処理して2時間以内に結果を提供する完全自動システムです。ナノプレート方式はサンプル調製のためのフロントエンド自動化に適しており、ハンズオン時間のさらなる削減をもたらします。

QIAcuity Nanoplateを使ったdPCRにおける独立した2つのランで得られた結果の再現性から、データの正確性を実証

存在比が大きく変動するcDNAターゲットのマルチプレックス解析の実現可能性を確証

Service Plans

QIAcuity One 2plex Full Agreement

Cat. No. / ID: 9245362

QIAcuity One 2Plex の派遣費、技術料および交換部品代を含む1年間の修理費用をカバー。契約期間内に1回の定期点検が付帯。対応時間2日以内

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QIAcuity One Preventive Subscription

Cat. No. / ID: 9245355

1回の予防保守（Preventive Maintenance, PM）点検訪問に対応します。交通費、人件費、予防保守部品費が含まれています。サブスクリプション期間における出張修理費、およびGMP/GLPに準拠した訪問報告書の作成費については10%減額します。

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QIAcuity Four Preventive Subscription

Cat. No. / ID: 9245356

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QIAcuity Eight Preventive Subscription

Cat. No. / ID: 9245357

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QIAcuity One 2plex Core Agreement

Cat. No. / ID: 9245358

年1回の出張修理と予防保守点検訪問に対応します。交通費、人件費、補修部品費が含まれています。受付から5営業日以内に対応します。契約期間における追加の出張修理費については10%減額します。

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QIAcuity One 5plex Core Agreement

Cat. No. / ID: 9245359

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QIAcuity Four Core Agreement

Cat. No. / ID: 9245360

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QIAcuity Eight Core Agreement

Cat. No. / ID: 9245361

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QIAcuity One 5plex Full Agreement

Cat. No. / ID: 9245363

QIAcuity One 5Plex の派遣費、技術料および交換部品代を含む1年間の修理費用をカバー。契約期間内に1回の定期点検が付帯。対応時間2日以内

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QIAcuity Four Full Agreement

Cat. No. / ID: 9245364

QIAcuity Four の派遣費、技術料および交換部品代を含む1年間の修理費用をカバー。契約期間内に1回の定期点検が付帯。対応時間2日以内

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QIAcuity Eight Full Agreement

Cat. No. / ID: 9245365

QIAcuity Eight の派遣費、技術料および交換部品代を含む1年間の修理費用をカバー。契約期間内に1回の定期点検が付帯。対応時間2日以内

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QIAcuity One Installation & Training

Cat. No. / ID: 9245352

装置ハードウェアの設置とシステムソフトウェアのインストールを実施します。装置の操作の実践体験と研究室スタッフの資格認定に役立つアプリケーショントレーニングを兼ね備えています。人件費、交通費、設置キット（該当する場合）が含まれています。

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QIAcuity Four Installation & Training

Cat. No. / ID: 9245353

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QIAcuity Eight Installation & Training

Cat. No. / ID: 9245354

このサービスはインストールや設置、ならびにお客様がQIAGENプラットフォームを使用したハードウェア、ソフトウェア、アプリケーションの実践体験を得るお手伝いをし、研究室スタッフが専門的な資格認定を取得する際に役立つアプリケーショントレーニングを兼ね備えています。

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Resources

For Version 2.0

Version 1.1.3

For Version 2

Version 4.0

The Volume Precision Factor (VPF) offers a unique feature to secure precision of concentration results obtained from a QIAcuity dPCR run.
In general, Nanoplates provide partitions of fixed sizes that enable a very precise way of sample concentration calculation. Potential variation of partition sizes in Nanoplate batches, caused by different microstructure molding forms, can be addressed by applying the batch specific VPF. Furthermore, the VPF includes well-specific volume information and therefore further increases precision of concentration calculation in each well of the Nanoplates.

After downloading and updating the VPF file within the QIAcuity Software Suite, the VPF is applied automatically to the analysis of a corresponding Nanoplate batch. The VPF file includes information from all available microstructure molding forms and connected Nanoplate batches. It will be stored on the PC where the QIAcuity Software Suite is installed.

Required QIAcuity Software Suite version: Version 1.2 or higher.

Version 2.1

Beside of usability improvements and bug fixes, the new features address GMP requirements by offering additional support for 21 CFR part 11 compliance, copy number variation (CNV) analysis, support of future Nanoplates, and improved 1D scatterplot views.

For more information, please refer to the Release Note: QIAcuity Software Suite, Version 2.1

SHA1 checksum: 6D51E273B26FA7D7E0EDA843CB3DCD74B38FA5B1

Version 2.0

The QIAcuity Control Software is an integral part of the QIAcuity instrument. It offers a GUI (graphical user interface) for basic functionalities such as plate setup, changing the order of plates to be processed, and monitoring the status of runs in real time. After a run is completed, the data are stored on the instrument’s memory and are sent to the connected QIAcuity Software Suite for analysis

Besides system stability and performance reliability improvements, this new version of the control software offers advanced user management and audit trail functionalities. Together with the QIAcuity Software Suite 2.0, the QIAcuity systems now support 21 CFR Part 11 regulations for users working under GMP.

Detailed information is available in the Release Note, which can also be downloaded under section “Operating Software”.

Note: Upgrading the QIAcuity Control Software to version 2.0 will require upgrading the QIAcuity Software Suite to version 2.0.

Important: Please follow the instructions provided in the QIAcuity User Manual for software version 2.0. It is strongly recommended to update the QIAcuity Software Suite first before proceeding with the QIAcuity Control Software!

Version 1.0

The QIAcuity Control Software is an integral part of the QIAcuity instrument. It offers a GUI (graphical user interface) for basic functionalities such as plate setup, changing the order of plates to be processed, and monitoring the status of runs in real time. After a run is completed, the data are stored on the instrument’s memory and are sent to the connected QIAcuity Software Suite for analysis.

This new version of the control software includes system stability and performance reliability improvements. We highly recommend to update the control software of your QIAcuity instrument.

Note: The installation requires approximately 90 minutes.

Version 2.0

The QIAcuity Software Suite 2.0 is designed to be installed on a Windows PC that is connected to one or more QIAcuity instruments. The QIAcuity Software Suite enables the user to set up plates, analyze results, and monitor the status of runs in real time. For this configuration, the QIAcuity instrument needs to be connected to a network through Ethernet. Alternatively, a direct cable connection between the QIAcuity and the notebook where the QIAcuity Software Suite is running needs to be established. When connected to a network, up to 10 users may access the QIAcuity Software Suite via a browser installed on the client PC (Windows or Mac).

This new software version now supports 21 CFR Part 11 regulations for users working under GMP. This includes the following:

-Advanced User Management
-Audit Trail
-Signatures

Additional changes compared with the previous software version include the following:

-Usability improvements
-Improved dust detection
-Hyperwell expansion to CNV and GEX analysis
-CSV export functionality of multiple positive partitions for linkage analysis, drop-off assays, etc.
-Raw data export

Detailed information is available in the Release Note, which can also be downloaded under section “Operating Software”.

Important: A direct update of the Software Suite from version 1.1.3 to 2.0.20 is not possible. Please follow the instructions in the User Manual on how to update to version 1.2.18 first before updating the Software Suite to version 2.0.20.

Before you start upgrading to version 1.1.3, please make sure that all plates are imported and visible in the plate overview of the QIAcuity Software Suite.

Caution: Not following the instructions may result in a loss of your previous plate data!

Please contact QIAGEN Technical Services if you are unsure and require technical support.

SHA1 checksum: C60C4EEC349DF1C037796016D32A93EEC19B54A0

Version 2.1

Beside of improvements and bug fixes, the new features mainly address the GMP requirements by offering additional support for 21 CFR part 11 compliance and get the software ready for future Nanoplate support.

For more information, please refer to the Release Note: QIAcuity Instrument Control Software, Version 2.1.

SHA1 checksum: 0DD41A8DE46D10E81F538ACB71AC184170D39FE0

Version 1.2

The QIAcuity Software Suite 1.2 is designed to be installed on a Windows PC that is connected to one or more QIAcuity instruments. The QIAcuity Software Suite enables the user to set up plates, analyze results, and monitor the status of runs in real time. For this configuration, the QIAcuity instrument needs to be connected to a network through Ethernet. Alternatively, a direct cable connection between the QIAcuity and the notebook where the QIAcuity Software Suite is running needs to be established. When connected to a network, up to 10 users may access the QIAcuity Software Suite via a browser installed on the client PC (Windows or Mac).

The following browsers are supported in the QIAcuity Software Suite:

-Mozilla Firefox (version 64.0.2 or higher)
-Microsoft Edge (version 44.17763.1.0 or higher)
-Google Chrome (version 71.0.3578.98 or higher)

The new QIAcuity Software Suite 1.2 offers a functionality that enables users of the QIAcuity Software 1.1.3 to upgrade to the new version while keeping the library of previously stored plate runs.

Note: If you have exported plates from QIAcuity Software Suite 1.1.3 that you would like to import and use in QIAcuity Software Suite 1.2, you will need to import the plates before upgrading from version 1.1.3 to version 1.2. You may then export the plates again. Future software version starting from QIAcuity Software Suite 2.0 will facilitate import of plates from previous QIAcuity Software Suite versions.

The new improvements are as follows:

-Support for the Nanoplate 8.5k 24-well
-Hyperwell functionality to combine several wells to one combined well for analysis
-Automated plate archiving functionality
-Functionality to show the number of single/double positives in 2D scatterplots
-VPF (Volume Precision Factor) to further improve concentration calculation (see related resources)
-Additional improvements for stabilization and troubleshooting

For Version 1.2

Version 2.0

-Advanced User Management
-Audit Trail
-Signatures

Additional changes compared with the previous software version include the following:

SHA1 checksum: C60C4EEC349DF1C037796016D32A93EEC19B54A0

For Version 1.0

This study tested a workflow for quantitation and qualification of AAV samples using a duplex assay on the QIAcuity dPCR instrument targeting both an insert (GFP) and the viral backbone (AAV2-ITR). With very low intra-assay and inter-assay CVs <6.5%, we demonstrate one of the main benefits of dPCR: reproducibility.

Here we report the use of saliva samples in combination with dPCR as a suitable alternative to screen for individuals infected with SARS-CoV-2.

The QIAgility instrument is a liquid handler designed for automating PCR setup. For compatibility with the QIAcuity, we developed an adapter to secure up to two nanoplates onto the deck of the QIAgility. Using the QIAgility software, we have optimized a protocol that works for all nanoplate types and QIAcuity applications. Here we report the performance of a front end automated QIAgility dPCR nanoplate setup procedure for use with the QIAcuity dPCR system.

The QIAcuity digital PCR system combined with the QIAcuity One-Step Viral RT-PCR Kit enables precise detection and quantitation of vector-borne viruses in mosquitoes. The results presented in this comparison study showed that digital PCR is a powerful tool for absolute quantitation of low abundant targets and is a more reliable detection method than qPCR. Multiplexing allows detection and quantitation of multiple targets in a single reaction more efficiently by increasing sample throughput at a reduced cost per target.

The goal of this work was to compare performance of quantitative PCR (qPCR) and digital PCR (dPCR) in the quantification of gene expression and Wolbachia abundances in Nasonia parasitoid wasps.

Here we provide an integrated rAAV genome titer method using the QIAcuity Digital PCR (dPCR) System with detailed parameters for high assay performance. Using this optimized method for pre-PCR handling of in-process rAAV samples, the results demonstrated that QIAcuity dPCR system generates the same level of accuracy and precision as the current gold standard ddPCR system but with much faster sample-to-result times (2 hours vs 7 hours) and higher overall throughput and scalability.

Here we compared the performance of qPCR and the nanoplate dPCR techniques. The digital PCR method on the QIAcuity significantly improved precision when measuring copy number states and sensitivity of mutation detection through absolute quantification and reduced standard error. This is advantageous in various applications, including copy number variation analysis, small fold-change and rare mutation detection.

Here we demonstrate how to optimize your assays on a microfluidic nanoplate-based digital PCR system, the QIAcuity, and provide recommendations for a seamless transfer. Moreover, the QIAcuity dPCR workflow is very similar to qPCR.

Fast. Scalable. Reliable.

A versatile workflow for the detection of low-abundance microbes

Updating QIAcuity Software Suite 1.1.3 and Control Software 0.5.2.18 to QIAcuity Software 2.0

Network connection of QIAcuity instruments

For QIAcuity User Manual – Updated QIAcuity Nanoplate Partition Volumes

User manual for QIAcuity instruments and QIAcuity Software 2.1

User manual for QIAcuity instruments and QIAcuity Software 1.2

For use with QIAcuity systems

July 2021

For QIAcuity Software Suite 1.2: Updating VPF for archived or exported plates

User manual for QIAcuity instruments and QIAcuity Software 2.0

A versatile workflow for the detection of low-abundance microbes

Fast. Scalable. Reliable.

Version 2.1

For more information, please refer to the Release Note: QIAcuity Instrument Control Software, Version 2.1.

SHA1 checksum: 0DD41A8DE46D10E81F538ACB71AC184170D39FE0

Version 1.1.3

Version 1.2

-Mozilla Firefox (version 64.0.2 or higher)
-Microsoft Edge (version 44.17763.1.0 or higher)
-Google Chrome (version 71.0.3578.98 or higher)

Version 2.0

-Advanced User Management
-Audit Trail
-Signatures

Additional changes compared with the previous software version include the following:

SHA1 checksum: C60C4EEC349DF1C037796016D32A93EEC19B54A0

Version 2.1

For more information, please refer to the Release Note: QIAcuity Software Suite, Version 2.1

SHA1 checksum: 6D51E273B26FA7D7E0EDA843CB3DCD74B38FA5B1

For Version 1.0

For Version 1.2

For Version 2.0

For Version 2

Version 2.1

Network connection of QIAcuity instruments

For QIAcuity Software Suite 1.2: Updating VPF for archived or exported plates

For QIAcuity User Manual – Updated QIAcuity Nanoplate Partition Volumes

Updating QIAcuity Software Suite 1.1.3 and Control Software 0.5.2.18 to QIAcuity Software 2.0

For use with QIAcuity systems

July 2021

User manual for QIAcuity instruments and QIAcuity Software 1.2

User manual for QIAcuity instruments and QIAcuity Software 2.0

User manual for QIAcuity instruments and QIAcuity Software 2.1

FAQ

The plate is designed for a single use run. For example, even if only 30 samples are loaded into the 96-well plate, a whole plate will be sealed by the roller. It can't be unsealed and used for another run. The QIAcuity Software won’t allow to set up a separate experiment for the same nanoplate to avoid that previously processed plates being not partitioned a second time.

3781

The QIAcuity Probe PCR Kit should be stored immediately upon receipt at –30 to –15°C in a constant-temperature freezer and protected from light. The QIAcuity Probe PCR master mix can also be stored protected from light at 2–8°C. Components are stable for 12 months, unless otherwise indicated on the label.
The QIAcuity EG PCR Kit should be stored immediately upon receipt at –30 to –15°C in a constant-temperature freezer and protected from light. The QIAcuity EG PCR master mix can also be stored protected from light at 2–8°C. Components are stable for 6 months, unless otherwise indicated on the label.
The QIAcuity Nanoplates does not have expiry date and are stable for at least 1 year when stored at RT.

FAQ ID - 9214

QIAGEN dPCR assays (such as dPCR LNA Mutation Assays) can be found on https://geneglobe.qiagen.com/.

FAQ ID - 9207

A standard PCR plate is required to set up dPCR reaction before transferring it to the nanoplate to ensure a proper mixing of the reaction mix before partitioning.

3774

The up-to-date VPF can be downloaded in the resource section of the QIAcuity product webpage. The VPF is compatible with the QIAcuity Software Suite 1.2 and higher. The latest VPF file contains all factors for all existing nanoplate batches. If a nanoplate from a new nanoplate batch is run and the latest version of the VPF was not installed, the software will recognize this and will give a message to install the latest VPF file. Please note that Applying the VPF file cannot be reversed.

FAQ ID - 9206

The VPF provides a set of well-specific and molding form-specific factors used to specify the exact reaction volume of a nanoplate, thus increasing the concentration calculation of each well.

FAQ ID - 9218

QIAGEN dPCR assays (such as dPCR LNA Mutation Assays) can be found on https://geneglobe.qiagen.com/.

3773

This is not needed. The QIAcuity is equipped with a flexible power supply technology and operates within a range of 100–240V AC, 50/60 Hz, 1500 VA (max).

3761

If you had run a nanoplate for which the installed VPF misses the specific factor, the software will notify you. If you then analyze without the specific VPF, the impact depends on the variation of the partition volume of the new Nanoplate batch compared to the latest. Typically this variation is ±6–7% (approx. 5% CV over the entire plate). The analysis may be repeated after updating the VPF file. After installing the latest VPF and re-analysis of the run, a copy of the plate is generated in the QIAcuity Software Suite including the new results.

3769

QIAGEN master mixes are optimized for nanoplate microfluidics and are recommended to be used with our dPCR system. They also include an optimized reference dye required for proper analysis.

3777

The QIAcuity reads emitted fluorescence from the bottom of the plate, which is covered with a foil. For best results, keep the foil clean and avoid damages such as scratches. Also, keep the barcode on the side of the plate clean and intact. Ensure that you wear gloves when working with a plate and do not apply force to it. For a safe handling of the plate, please place the plate into a nanoplate tray.

FAQ ID - 9202

QIAGEN master mixes are optimized for nanoplate microfluidics and are recommended to be used with our dPCR system. They also include an optimized reference dye required for proper analysis.

FAQ ID - 9211

QIAcuity does not support temperature gradient in PCR cycling profile. However, optimization of a dPCR assay can be done by qPCR on a gradient cycler using the dPCR master mix and then transferred from qPCR to dPCR.

3783

QIAGEN offers a complete range of nucleic acid purification systems. These include QIAprep kits for purification of plasmid DNA, QIAamp and DNeasy kits for purification of genomic DNA, RNeasy kits for purification of total RNA, and the PAXgene Blood RNA System for stabilization and purification of RNA from blood. Phenol and other contaminants can be efficiently removed from crude RNA preps using the RNeasy MinElute Cleanup Kit to clean up and concentrate RNA for sensitive assays. Details about QIAGEN kits for nucleic acid purification can be found at www.qiagen.com.

FAQ ID - 9213

The QIAcuity instrument software does not allow to read and process a plate without seal. If you would like to perform a dry run please use sealed plate and set up this plate in the QIAcuity Software Suite.

FAQ ID - 9201

dPCR master mix can be used in qPCR to optimize sample concentration and/or primer/probe concentration prior to assay transfer from qPCR to dPCR.

FAQ ID - 9209

This is not needed. The QIAcuity is equipped with a flexible power supply technology and operates within a range of 100–240V AC, 50/60 Hz, 1500 VA (max).

FAQ ID - 9195

Results are stored as part of the plates within the QIAcuity Software Suite. In version 1.1.2, plates can be exported to another file location, for example, an external HDD, and imported again if needed. From version 1.2 onwards, plates can also be archived automatically. To do so, an archive destination has to be defined. Additional information can be found in the user manual.

FAQ ID - 9198

QIAGEN offers a complete range of nucleic acid purification systems. These include QIAprep kits for purification of plasmid DNA, QIAamp, and DNeasy kits for purification of genomic DNA, RNeasy kits for purification of total RNA, and the PAXgene Blood RNA System for stabilization and purification of RNA from blood. Phenol and other contaminants can be efficiently removed from crude RNA preps using the RNeasy MinElute Cleanup Kit to clean up and concentrate RNA for sensitive assays. Details about QIAGEN kits for nucleic acid purification can be found at www.qiagen.com.

3779

If you had analyzed your nanoplates without VFP, the impact depends on the variation of the partition volume of the new nanoplate batch compared to the latest. The VPF reduces the CV from approximately 5% to 2%. We recommend to reanalyze results in case the data originated from different wells (e.g., copy number variations or gene expression data sets for which the reference sample was measured in a different well). Results obtained across different plates should also be r-analyzed. A reanalysis is not required for assay data that were analyzed within the same well (e.g., mutation rate determination using two channels within the same well).

3771

In dPCR we measure the absolute concentration of targets at endpoint reaction. Concentrations of unknowns can be determined based on dPCR results observed (number of negatives, number of positives, and partition volume analyzed).

3786

Both software are designed to be upgraded by users. The user manual includes instructions on how to perform the upgrades; instructions for the QIAcuity Suite Software upgrade can be found at page 38 and instructions for the CSW upgrade on page 67 (QIAcuity User Manual, 03/2021).

FAQ ID - 9196

If you had analyzed your nanoplates without VFP, the impact depends on the variation of the partition volume of the new nanoplate batch compared to the latest. The VPF reduces the CV from approximately 5% to 2%. We recommend to re-analyze results in case the data originated from different wells (e.g., copy number variations or gene expression data sets for which the reference sample was measured in a different well). Results obtained across different plates should also be re-analyzed. A re-analysis is not required for assay data that were analyzed within the same well (e.g., mutation rate determination using two channels within the same well).

FAQ ID - 9205

In general, nanoplates provide partitions of fixed sizes that enable a very precise way of sample concentration calculation. If a new molding form is used for nanoplate manufacturing, potential variation of partition sizes can be addressed by applying the molding form-specific VPF. Thus, each time a new molding form is used, a new VPF is created and made available. Currently, the VPF is updated once every 3–6 months.

3785

FAQ ID - 9219

Nanoplate 26K 24-well: 40 μl
Nanoplate 8.5K 24-well:12 μl
Nanoplate 8.5K 96-well: 12 μl

FAQ ID - 9216

The fluorescent signal in the reference channel is measured to determine the number of valid partitions in a well. In addition, differences in the signal intensities between partitions are normalized and the fluorescent signals in the target channels are corrected accordingly.

FAQ ID - 9210

In dPCR, we measure the absolute concentration of targets at endpoint reaction. Concentrations of unknowns can be determined based on dPCR results observed (number of negatives, number of positives, and partition volume analyzed).

FAQ ID - 9220

3776

The VPF provides a set of well-specific and molding form-specific factors used to specify the exact reaction volume of a nanoplate, thus increasing the concentration calculation of each well.

3784

Yes, the report includes a notification if the matching VPF was missing and, therefore, not applied to the analysis. If the matching VPF was applied there is no notification on the report.

3770

3780

Nanoplate 26K 24-well: 40 μl
Nanoplate 8.5K 24-well:12 μl
Nanoplate 8.5K 96-well: 12 μl

3782

All DNA samples used in reaction mixes should show similar quality and quantity, which can easily be assessed using UV spectrophotometry. DNA samples with an average length of ≥20 kb (e.g., genomic DNA purified via spin column with silica membrane) should be fragmented by restriction digestion before partitioning. Enzymatic fragmentation of larger DNA ensures an even distribution of template throughout the QIAcuity Nanoplate, which in turn leads to an accurate and precise quantification.

3778

The instrument software GUI shows error codes including a description and information how to resolve the error. The instrument touchscreen shows an alarm icon in the upper right corner that turns red in case of an instrument failure. Accessing the System Status in the Tool tab allows users to clear errors. Rebooting of the instrument is required to complete the removal of the error. Please do not skip this step. You may always contact QIAGEN Technical Services in case of any question.

3763

FAQ ID - 9215

FAQ ID - 9203

Yes, the report includes a notification if the matching VPF was missing and, therefore, not applied to the analysis. If the matching VPF was applied there is no notification on the report.

FAQ ID - 9204

All DNA samples used in reaction mixes should show similar quality and quantity, which can easily be assessed using UV spectrophotometry. DNA samples with an average length of ≥20 kb (e.g., genomic DNA purified via spin column with silica membrane) should be fragmented by restriction digestion before partitioning. Enzymatic fragmentation of larger DNA ensures an even distribution of template throughout the QIAcuity nanoplate, which in turn leads to an accurate and precise quantification.

FAQ ID - 9212

3767

dPCR master mix can be used in qPCR to optimize sample concentration and/or primer/probe concentration prior to assay transfer from qPCR to dPCR.

3775

The user manual contains instructions on how to perform a regular cleaning and decontamination, and how to replace air filters on the QIAcuity instruments. A regular maintenance reduces the dust in the instrument and therefore minimizes the presence of dust particles on the nanoplate, which might interfere with the plate analysis.

3765

3772

3768

FAQ ID - 9197

No. The QIAcuity platform introduces four variations: QIAcuity One 2plex, QIAcuity One 5plex, QIAcuity Four (5plex), and QIAcuity Eight (5plex). All of them have fix channel combination.

FAQ ID - 9200

No. The QIAcuity platform introduces four variations: QIAcuity One 2plex, QIAcuity One 5plex, QIAcuity Four (5plex), and QIAcuity Eight (5plex). All of them have fix channel combination.

3766

A standard PCR plate is required to set up dPCR reaction before transferring it to the nanoplate to ensure a proper mixing of the reaction mix before partitioning.

FAQ ID - 9208

FAQ ID - 9217

3764

3762

The user manual contains instructions on how to perform a regular cleaning, decontamination, and how to replace air filters on the QIAcuity instruments. A regular maintenance reduces the dust in the instrument and therefore minimizes the presence of dust particles on the nanoplate, which might interfere with the plate analysis.

FAQ ID - 9199