QIAcuity Digital PCR System

나노플레이트 기반 디지털 PCR 애플리케이션

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QIAcuity One, 2plex Device

Cat. No. / ID: 911001

최대 2가지 형광 염료 검출용 1개 플레이트 디지털 PCR 기기, 롤러, USB 플래시 메모리 및 QIAcuity Software Suite: 1회의 예방 유지보수 방문 포함. 또한 공임, 출장 및 부품 1년 보증 포함.

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Format

One

Four

Eight

Multi-channel detection

2plex

5plex

Service Options

Device

System

System FUL-2

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QIAcuity는 분자생물학 분야에 사용하기 위한 것입니다. 이 제품은 질병의 진단, 예방, 또는 치료 목적으로 사용할 수 없습니다. 따라서 임상 사용을 위한 제품의 성능 특징(즉, 진단, 예후, 치료 또는 혈액은행)은 알려져 있지 않습니다.

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Features

완전 통합 시스템
확장 가능 형식(1개, 4개, 8개 플레이트 기기)
고급 멀티플렉싱 기능(최대 5 플렉스)
유연성 있는 샘플 처리량
약 2시간 내에 포괄적인 결과 산출

Product Details

QIAcuity Digital PCR System은 돌연변이 검출, 복제 수 변이(CNV), 유전자 발현 연구, 유전자 편집 분석 등에 대한 정밀하고 멀티플렉싱된 정량화 결과를 제공하도록 설계되었습니다. 이 나노플레이트 기반 시스템은 분획, 열순환(thermocycling) 및 이미징의 표준 dPCR 워크플로우를 최소한의 직접 작업 시간으로 자동화된 플랫폼에 완벽하게 통합합니다.

이 시스템은 QIAcuity Nanoplates 및 액세서리와 함께 사용합니다.

가상 데모를 통해 QIAcuity에 대해 더 자세히 알아보세요.

Performance

QIAcuity Digital PCR System은 모든 실험실에서 절대 정량화를 합리적인 비용으로 할 수 있게 합니다. 워크어웨이 자동화는 최소한의 작업 시간으로 분획, 열순환(thermocycling) 및 이미징의 전체 디지털 PCR 워크플로우를 하나의 기기로 통합하여 간소화합니다. 또한 현재 qPCR 분석 항목을 QIAcuity Digital PCR System에 쉽게 적용할 수 있습니다. qPCR에서 전환할 때 플레이트 사용 방식에는 변화가 필요 없으므로 약 2시간 내에 빠른 분석 항목 설정과 빠른 결과를 얻을 수 있습니다.

QIAcuity 기기 – 기능 및 사양

기능	QIAcuity One	QIAcuity Four	QIAcuity Eight
처리 플레이트	1	4	8
검출 채널(멀티플렉싱)	2 또는 5	5	5
서모사이클러	1	1	2
결과까지 걸리는 시간	약 2시간	첫 번째 플레이트는 약 2시간 그다음 플레이트당 ~60분	첫 번째 플레이트는 약 2시간 그다음 플레이트당 ~30분
처리량(작업일 하루에 처리하는 검체)	최대 384개(96웰) 최대 96개(24웰)	최대 672개(96웰) 최대 168개(24웰)	최대 1248개(96웰) 최대 312개(24웰)

Principle

피펫팅 및 로딩, 실험 실행, 결과 분석의 간단한 3단계로 약 2시간 내에 원하는 dPCR 결과를 얻을 수 있습니다.

나노플레이트에서의 dPCR 반응 원리는 여기에 설명되어 있습니다.

Procedure

qPCR 실험과 마찬가지로 샘플 준비에는 마스터 믹스, 프로브 및 프라이머를 96웰 또는 24웰 나노플레이트로 옮긴 다음 샘플을 추가하는 작업이 포함됩니다. 이 시스템은 분획, 열순환(thermocycling) 및 이미징을 하나의 완전 자동화 기기로 통합하여 사용자가 2시간 이내에 샘플에서부터 결과를 얻을 수 있습니다. Suite 소프트웨어에서 분석을 수행할 수 있으며, 표적 서열의 마이크로리터당 복제 농도뿐만 아니라 양성 검체나 NTC와 같은 품질 관리도 제공됩니다. 이 분석은 또한 동일한 LAN(근거리 통신망) 내의 원격 컴퓨터로 확장될 수 있습니다.

Applications

QIAcuity 기기는 QIAcuity Nanoplates 및 QIAcuity PCR 키트와 함께 다음을 포함한 디지털 PCR 애플리케이션을 지원합니다.

희귀 돌연변이 검출
복제 수 변이 분석
유전자 발현 분석
병원체 검출
유전형 분석
miRNA 연구

Software

기기와 함께 제공되고 별도의 컴퓨터에 설치된 QIAcuity Software Suite는 한 대의 기기에 직접 연결하거나 기존 LAN(근거리 통신망)을 사용하여 한 대 이상의 QIAcuity 기기를 제어합니다. QIAcuity Software Suite를 사용하여 디지털 PCR 실험, 샘플 및 반응 혼합물을 정의하고 Nanoplate에 배정하고 QIAcuity 기기로 전송할 수 있습니다. 실행 후 데이터를 분석하고 보고서를 만들고 외부 분석을 위해 데이터를 내보낼 수 있습니다. 이 소프트웨어는 반복되는 플레이트 레이아웃 또는 플레이트 실행 매개변수에 쉽게 액세스할 수 있도록 하는 여러 템플릿 기능을 제공하여 디지털 PCR 경험을 더욱 향상합니다.

LAN에 통합되면 컴퓨터가 LAN을 통해 클라이언트 역할을 하는 다른 컴퓨터에 액세스할 수 있는 서버로서 QIAcuity Software Suite 기능을 호스팅합니다. 이를 통해 여러 사용자가 여러 컴퓨터에 소프트웨어를 설치하거나 인터넷 연결을 통해 데이터에 액세스하고 교환할 필요 없이 다른 방이나 사무실에서 소프트웨어에 액세스하고 표준 브라우저를 통해 데이터를 분석할 수 있습니다.

Services

QIAGEN의 다양한 서비스 솔루션으로 기기를 관리하십시오. 귀하의 필요성에 맞는 특정 서비스 계약에 대해 알아보십시오.

Supporting data and figures

간단하고 신속한 플레이트 기반 워크플로우

분획, 서모사이클링 및 이미징이 모두 하나의 완전히 자동화된 기기에 통합되어 2시간 이내에 결과를 제공합니다. 나노플레이트 형식은 샘플 준비를 위한 프런트 엔드 자동화가 가능하여 직접 실험 시간이 더욱 단축됩니다.

QIAcuity Nanoplate dPCR을 사용한 두 번의 독립적 실행에서 반복 가능한 결과로 데이터 정확도가 입증되었습니다.

매우 다양한 존재비의 cDNA 표적의 멀티플렉싱 가능성이 확인되었습니다.

Service Plans

QIAcuity One 2plex Full Agreement

Cat. No. / ID: 9245362

1년간 출장, 공임 및 부품을 포함한 현장 QIAcuity One 2plex 기기 수리. 영업일 기준 2일 대응 시간. 전체 계약 기간 동안 1회 예방 유지보수 포함.

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QIAcuity One Preventive Subscription

Cat. No. / ID: 9245355

출장, 공임 및 예비 부품을 포함한 1회 예방 유지보수(PM). 예방 서비스 가입 기간 동안 수리 서비스 10% 할인 및 GMP/GLP 준수 방문 보고서 문서화 포함.

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QIAcuity Four Preventive Subscription

Cat. No. / ID: 9245356

출장, 공임 및 예비 부품을 포함한 1회 예방 유지보수(PM). 예방 서비스 가입 기간 동안 수리 서비스 10% 할인 및 GMP/GLP 준수 방문 보고서 문서화 포함.

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QIAcuity Eight Preventive Subscription

Cat. No. / ID: 9245357

출장, 공임 및 예비 부품을 포함한 1회 예방 유지보수(PM). 예방 서비스 가입 기간 동안 수리 서비스 10% 할인 및 GMP/GLP 준수 방문 보고서 문서화 포함.

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QIAcuity One 2plex Core Agreement

Cat. No. / ID: 9245358

1년간 출장, 공임 및 예비 부품을 포함한 1회 현장 수리 서비스 및 1회 현장 예방 유지보수. 영업일 기준 5일 대응 시간. 계약 기간 동안 추가 수리 서비스에 대한 10% 할인 포함.

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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 2plex Basic Agreement

Cat. No. / ID: 9245401

1년간 출장, 공임 및 부품을 포함한 현장 QIAcuity One 2plex 기기 수리. 영업일 기준 5일 대응 시간. 기본 계약 기간 동안 1회 예방 유지보수 또는 점검 서비스 포함.

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QIAcuity One 5plex Basic Agreement

Cat. No. / ID: 9245402

1년간 출장, 공임 및 부품을 포함한 현장 QIAcuity One 5plex 기기 수리. 영업일 기준 5일 대응 시간. 기본 계약 기간 동안 1회 예방 유지보수 또는 점검 서비스 포함.

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QIAcuity Four Basic Agreement

Cat. No. / ID: 9245403

1년간 출장, 공임 및 부품을 포함한 현장 QIAcuity Four 기기 수리. 영업일 기준 5일 대응 시간. 기본 계약 기간 동안 1회 예방 유지보수 포함.

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QIAcuity Eight Basic Agreement

Cat. No. / ID: 9245404

1년간 출장, 공임 및 부품을 포함한 현장 QIAcuity Eight 기기 수리. 영업일 기준 5일 대응 시간. 기본 계약 기간 동안 1회 예방 유지보수 포함.

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QIAcuity One 5plex Full Agreement

Cat. No. / ID: 9245363

1년간 출장, 공임 및 부품을 포함한 현장 QIAcuity One 5plex 기기 수리. 영업일 기준 2일 대응 시간. 전체 계약 기간 동안 1회 예방 유지보수 포함.

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QIAcuity Four Full Agreement

Cat. No. / ID: 9245364

1년간 출장, 공임 및 부품을 포함한 현장 QIAcuity Four 기기 수리. 영업일 기준 2일 대응 시간. 전체 계약 기간 동안 1회 예방 유지보수 포함.

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QIAcuity Eight Full Agreement

Cat. No. / ID: 9245365

1년간 출장, 공임 및 부품을 포함한 현장 QIAcuity Eight 기기 수리. 영업일 기준 2일 대응 시간. 전체 계약 기간 동안 1회 예방 유지보수 포함.

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QIAcuity One Installation & Training

Cat. No. / ID: 9245352

기기 하드웨어 및 시스템 소프트웨어 설치. 기기 작동 및 실험실 직원에 대한 자격 부여와 관련된 실무 경험을 위한 애플리케이션 교육 포함. 공임비, 출장비 및 설치 키트(해당하는 경우) 포함.

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QIAcuity Four Installation & Training

Cat. No. / ID: 9245353

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QIAcuity Eight Installation & Training

Cat. No. / ID: 9245354

이 서비스는 애플리케이션 교육과 설치를 병합하여 고객이 QIAGEN 플랫폼을 이용하는 하드웨어, 소프트웨어, 애플리케이션을 직접 경험할 수 있도록 지원하며 실험실 직원이 전문 자격을 갖출 수 있도록 합니다.

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Resources

For Version 1.0

For Version 1.2

For Version 2.0

For Version 2

Version 2.1

Version 2.1.8

Version 2.2

Version 2.5

Version 2.5.0.0

Version 2.5.0.1

Network connection of QIAcuity instruments

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

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

For QIAcuity User Manual – Updated QIAcuity Nanoplate Partition Volumes

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

User manual for QIAcuity instruments and QIAcuity Software 2.1.8

User manual for QIAcuity instruments and QIAcuity Software 2.2

User manual for QIAcuity instruments and QIAcuity Software 2.5

QIAcuity backup and restore scripts for QIAcuity Software Suite versions 2.0, 2.1.7, 2.1.8, and 2.2

QIAcuity backup and restore scripts for QIAcuity Software Suite version 2.5 or higher

Version 2.5.0.1

The QIAcuity Software Suite enables the user to set up plates, analyze results, and monitor the status of runs in real time. This software is also used for the configuration of the system and provides access to the QIAcuity user management. The QIAcuity Software Suite is designed to be installed on a Windows PC that is connected to one or more QIAcuity instruments. 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 new version 2.5 of the QIAcuity Software Suite supports assay development by providing an essential temperature gradient functionality. It also offers a new feature that provides calculation of initial concentration of samples by using various dilution factors. In addition, concentration units may be converted into various pre-defined or user-defined units. The new software also offers an integrity value and concentration value per group for up to 5-plex added to the multiple occupancy CSV file export, for example, for the evaluation of AAV (adeno-associated virus) assays and for drop-off assays.

In this new version the initial loading time and the time for recalculation of 1D/2D scatterplot and for signal map image has been reduced, leading to a much faster performance.

Detailed information about the QIAcuity Software Suite version 2.5 is available in the Release Note, which can also be downloaded under section “Software Release Notes”.

Note: The latest Software Suite version 2.5 is only compatible with the latest Control Software (CSW) version 2.5. If only one software component is updated, no connection between the Software Suite and the CSW can be established.

Important: Please follow the instructions provided in the QIAcuity User Manual for software version 2.5 and in the Release Note.

It is strongly recommended to update the QIAcuity Software Suite first before proceeding with the QIAcuity Control Software!

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

SHA1 checksum: D1690226299A75E077FA37C420A970FC71D56CBF

Version 8.0

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.

Important note:

The Volume Precision Factor file version 8.0 is compatible with QIAcuity Software Suite version 2.1.8.23 or higher. All lower versions, namely 1.2.18, 2.0.20, 2.1.7.182, and 2.1.8.20, require a QIAcuity Software Suite patch to be installed prior the upload of VPF version 8.0. Please read the Release Note for QIAcuity Software Suite Volume Precision Factor (VPF) Patches for more information.

If you are using QIAcuity Software Suite versions older than 2.1.8.23, please consider updating your Software Suite and the Instrument Control Software (CSW) to the most recent version prior the upload of VPF version 8.0. After updating to the latest QIAcuity Software Suite software version, no patching is needed. If you are not able to update your QIAcuity Software Suite, please run the patch for your Software Suite version following the instructions provided in the Release Note for QIAcuity Software Suite Volume Precision Factor (VPF) Patches.

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.

The following QIAcuity Software Suite Volume Precision Factor (VPF) patches have been released to enable compatibility with VPF file version 6.0 or higher for QIAcuity Software Suite version 1.2.18. As the abovementioned QIAcuity Software Suite versions do not allow loading a VPF zip file including more than 20 different individual VPF files, a patch is needed to allow the software for loading VPF zip files version 6.0 or higher. This technical limitation is solved from QIAcuity Software Suite version 2.1.8.23 onwards.

If you are using the QIAcuity Software Suite version 1.2.18, please consider updating your Software Suite and the Instrument Control Software (CSW) to the most recent version. After updating to the latest software version, no patching is needed.

If you are not able to update your QIAcuity Software Suite, please install the QIAcuity Software Suite VPF patch for QIAcuity Software Suite version 1.2.18 before loading VPF file version 6 or higher. Additional information and instructions may be found on the Release Note: QIAcuity Software Suite Volume Precision Factor (VPF) Patches.

The following QIAcuity Software Suite Volume Precision Factor (VPF) patches have been released to enable compatibility with VPF file version 6.0 or higher for QIAcuity Software Suite versions 2.0.20. As the abovementioned QIAcuity Software Suite versions do not allow loading a VPF zip file including more than 20 different individual VPF files, a patch is needed to allow the software for loading VPF zip files version 6.0 or higher. This technical limitation is solved from QIAcuity Software Suite version 2.1.8.23 onwards.

If you are using the QIAcuity Software Suite version 2.0.20, please consider updating your Software Suite and the Instrument Control Software (CSW) to the most recent version. After updating to the latest software version, no patching is needed.

If you are not able to update your QIAcuity Software Suite, please install the QIAcuity Software Suite VPF patch for QIAcuity Software Suite version 2.0.20 before loading VPF file version 6 or higher. Additional information and instructions may be found on the Release Note: QIAcuity Software Suite Volume Precision Factor (VPF) Patches.

The following QIAcuity Software Suite Volume Precision Factor (VPF) patches have been released to enable compatibility with VPF file version 6.0 or higher for QIAcuity Software Suite version 2.1.7.182. As the abovementioned QIAcuity Software Suite versions do not allow loading a VPF zip file including more than 20 different individual VPF files, a patch is needed to allow the software for loading VPF zip files version 6.0 or higher. This technical limitation is solved from QIAcuity Software Suite version 2.1.8.23 onwards.

If you are using the QIAcuity Software Suite version 2.1.7.182, please consider updating your Software Suite and the Instrument Control Software (CSW) to the most recent version. After updating to the latest software version, no patching is needed.

If you are not able to update your QIAcuity Software Suite, please install the QIAcuity Software Suite VPF patch for QIAcuity Software Suite version 2.1.7.182 before loading VPF file version 6 or higher. Additional information and instructions may be found on the Release Note: QIAcuity Software Suite Volume Precision Factor (VPF) Patches.

The following QIAcuity Software Suite Volume Precision Factor (VPF) patches have been released to enable compatibility with VPF file version 6.0 or higher for QIAcuity Software Suite version 2.1.8.20. As the abovementioned QIAcuity Software Suite versions do not allow loading a VPF zip file including more than 20 different individual VPF files, a patch is needed to allow the software for loading VPF zip files version 6.0 or higher. This technical limitation is solved from QIAcuity Software Suite version 2.1.8.23 onwards.

If you are using the QIAcuity Software Suite version 2.1.8.20, please consider updating your Software Suite and the Instrument Control Software (CSW) to the most recent version. After updating to the latest software version, no patching is needed.

If you are not able to update your QIAcuity Software Suite, please install the QIAcuity Software Suite VPF patch for QIAcuity Software Suite version 2.1.8.20 before loading VPF file version 6 or higher. Additional information and instructions may be found on the Release Note: QIAcuity Software Suite Volume Precision Factor (VPF) Patches.

The QIAcuity backup and restore scripts are a standalone solution to backup all relevant user data of the QIAGEN Software Suite for disaster recovery and restore the data on a new or existing QIAGEN Software Suite installation. The following QIAcuity Software Suite versions are supported: 2.0, 2.1.7, 2.1.8, and 2.2. The backup can be conducted manually or automated by using of the Windows task scheduler.

Note: Windows Admin permission is needed to setup and perform an automated backup and for manual backup and restore.

Please read the QIAcuity Software Suite Backup and Restore Scripts document for more information and instructions how to use the scripts.

The QIAcuity backup and restore scripts are a standalone solution to backup all relevant user data of the QIAGEN Software Suite for disaster recovery and to restore the data on a new or existing QIAGEN Software Suite installation. The backup supports the QIAcuity Software Suite version 2.5 or higher and can be conducted manually or automatically by using of the Windows Task Scheduler.

Note: Windows Admin permission is needed to setup and perform an automated backup and for manual backup and restore.

For backup and restore of Software versions lower than 2.5, please refer to the QIAcuity Software Suite Backup and Restore Scripts for version 2.0, 2.1.7, 2.1.8 and 2.2.

Note: Windows Admin permission is needed to setup and perform an automated backup and for manual backup and restore.

Please read the QIAcuity Software Suite Backup and Restore Scripts document for more information and instructions how to use the scripts.

Version 2.5

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.

The new version 2.5 of the QIAcuity CSW offers a configurable auto logoff times which enables users to turn off or define logoff times per instrument. In addition the new version supports assay development by providing an essential temperature gradient functionality.

Furthermore improvements were implemented, for example, for the accuracy of time estimation for various software steps.

Detailed information about the QIAcuity Control Software version 2.5 is available in the Release Note, which can also be downloaded under section Software Release Notes.

Note: The latest CSW version 2.5 is only compatible with the Software Suite version 2.5. If only one software component is updated, no connection between the Software Suite and the CSW can be established.

Important: Please follow the instructions provided in the QIAcuity User Manual for software version 2.5 and in the Release Note.

It is strongly recommended to update the QIAcuity Software Suite first before proceeding with the QIAcuity Control Software!

Note: After clicking reboot during CSW upgrade or change of Suite connection, the login screen may appear for short period. Please ignore it and wait for the QIAcuity instrument to shut down and restart itself.

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

Here, we present a highly efficient, high-throughput workflow that combines two technologies, cellenONE and QIAcuity Digital PCR, to accurately analyze miRNAs in well-defined individual cells and populations of cells.

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 present a workflow that combines two technologies, cellenONE and QIAcuity Digital PCR, which accelerate and streamline high-throughput analyses of target copy numbers in cultured cells. The workflow starts with detecting and sorting defined populations of cells as well as individual cells using cellenONE, followed by multiplexing dPCR on the QIAcuity platform. Copy number variations of target regions are then analyzed using the QIAcuity Software Suite, providing an intuitive and fast interpretation of results.

With the cellenONE-QIAcuity workflow, we achieved high-throughput absolute quantification of low-abundance targets at the single-cell level. cellenONE’s single-cell isolation platform enabled real-time and 100% accurate single-cell isolation without compromising viability and transcript expression. This allowed us to use the exact number of intact cells for RT-dPCR reactions. Moreover, the elimination of the RNA purification step significantly reduced hands-on time, and QIAcuity’s probe-based chemistry allowed the multiplexing of up to five targets in a one-step RT-dPCR format with no or minimal optimization.

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.

Q-Solution is an innovative and versatile PCR additive that works in part by lowering the melting temperature of DNA. Adjusting template melting properties can be useful when interrogating DNA with high-GC content or with complex secondary structures, such as adeno-associated virus (AAV) genomes. Q-Solution has additional properties that can be used to mitigate the carryover of various PCR inhibitors. This application note will highlight a few examples of how the Q-Solution Kit can be used in QIAcuity workflows.

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.

A versatile workflow for the detection of low-abundance microbes

Fast. Scalable. Reliable.

Version 2.1.8

The new version 2.1.8 of the QIAcuity software (QIAcuity Software Suite version 2.1.8 and QIAcuity CSW version 2.1.8.) offers bug fixes for repeated network connection issues and improved error handling. In addition, it fixes issues seen with latest versions of Microsoft Edge and Google Chrome browsers as well as issues seen with read-only plates after a software update.

Detailed information about the QIAcuity Software Suite version 2.1.8 is available in the Release Note, which can also be downloaded under section “Software Release Notes”.

Important: Please ensure that updating to QIAcuity Software Suite version 2.1.8 is performed by the same Windows admin user using the same Windows login name that installed the previous QIAcuity Software Suite version. In case you cannot use the same Windows login name or the software update resulted in the browser error message "Can't reach this page" please contact our Technical Services for additional instructions.

Note: The latest Software Suite version 2.1.8 is only compatible with the latest Control Software (CSW) version 2.1.8. If only one software component is updated, no connection between the Software Suite and the CSW can be established.

Important: Please follow the instructions provided in the QIAcuity User Manual for software version 2.1 and in the Release Note. It is strongly recommended to update the QIAcuity Software Suite first before proceeding with the QIAcuity Control Software!

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

SHA1 checksum: DE2416D926BB98D82A332E7B25EF66728F120DDA

Version 2.2

The new version 2.2 of the QIAcuity software (QIAcuity Software Suite v 2.2 and QIAcuity CSW v 2.2.) offers improvements for users working under GMP by adding a user ID validation during the report signing and the addition of timezone offset stamp for audit trail entries and for result report data. Furthermore, the addition of a standard deviation and coefficient of variance calculation in percentage of mean concentration calculation for replicates where implemented. In addition, the instrument camera stability was improved and an internal validation method was implemented.

Detailed information about the QIAcuity Software Suite version 2.2 is available in the Release Note, which can also be downloaded under section “Software Release Notes”.

Note: The latest Software Suite version 2.2 is only compatible with the latest Control Software (CSW) version 2.2. If only one software component is updated, no connection between the Software Suite and the CSW can be established.

Important: Please follow the instructions provided in the QIAcuity User Manual for software version 2.2 and in the Release Note.

It is strongly recommended to update the QIAcuity Software Suite first before proceeding with the QIAcuity Control Software.

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

SHA1 checksum: 63A1689F2EB557809F418D159DA438FDC8B327A3

Version 2.1.8

The QIAcuity Control Software (CSW) 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.

The new version 2.1.8 of the QIAcuity software (QIAcuity Software Suite version 2.1.8 and QIAcuity CSW version 2.1.8) offers bug fixes for repeated network connection issues and improved error handling. In addition, it fixes issues seen with latest versions of Microsoft Edge and Google Chrome browsers as well as issues seen with read-only plates after a software update.

Detailed information about the QIAcuity Control Software version 2.1.8 is available in the Release Note, which can also be downloaded under section “Software Release Notes”.

Note: The latest CSW version 2.1.8 is only compatible with the latest Software Suite version 2.1.8. If only one software component is updated, no connection between the Software Suite and the CSW can be established.

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

SHA1 checksum: D3982B419D7C4CF39FBDDDAA4C0D351B39398278

Version 1.2

A newer version of the Software Suite is available. Please use this version for the update of older plates if required.

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

Version 2.2

The new Version 2.2 of the QIAcuity software (QIAcuity Software Suite v 2.2 and QIAcuity CSW v 2.2.) offers improvements for users working under GMP by adding a user ID validation during the report signing and the addition of timezone offset stamp for audit trail entries and for result report data. Furthermore, the addition of a standard deviation and coefficient of variance calculation in percentage of mean concentration calculation for replicates where implemented. In addition, the instrument camera stability was improved and an internal validation method was implemented.

Detailed information about the QIAcuity Control Software version 2.2 is available in the Release Note, which can also be downloaded under section “Software Release Notes”.

Note: The latest CSW version 2.2 is only compatible with the Software Suite version 2.2. If only one software component is updated, no connection between the Software Suite and the CSW can be established.

Important: Please follow the instructions provided in the QIAcuity User Manual for software version 2.2 and in the Release Note.

It is strongly recommended to update the QIAcuity Software Suite first before proceeding with the QIAcuity Control Software.

Note: After clicking reboot during CSW upgrade or change of Suite connection, login screen may appear for short period. Please ignore it and wait for the QIAcuity instrument to shut down and restart itself.

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

SHA1 checksum: 3892D7A434A8F072A15008C76EB088BB78F1C255

Compliance with 21 CFR Part 11 involves both technical (i.e., hardware and software) and procedural requirements. This document explains how the QIAcuity system contributes to fulfilling those technical requirements.

A new temperature gradient functionality to determine the optimal primer annealing temperature of a digital PCR assay

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

An essential temperature gradient functionality was introduced with software version 2.5. When updating older software versions to 2.5, each QIAcuity instrument will offer the temperature gradient and may be used to find the best cycling temperature for new dPCR assays. When running a QIAcuity Four or a QIAcuity Eight, all plates may have their own temperature profile, including the option for a temperature gradient.

FAQ ID - 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

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.

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