High Throughput Real-Time PCR (qPCR) for AAC Genomics Core Lab

AAC 基因组学核心实验室的高通量实时 PCR (qPCR)

• 批准号: RTI-2020-00643
• 负责人: VanDerKraak, Glen
• 金额: $ 10.65万
• 依托单位: University of Guelph
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2019
• 资助国家: 加拿大
• 起止时间: 2019-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=693609
• 关键词: Throughput; Real; Time; PCR; qPCR

The major users of the Advanced Analysis Centre (AAC) Genomics Facility, which offers core molecular biology technology support services and training to the entire UofG campus, are requesting funding for a QuantStudio 7 Pro real-time PCR system to replace existing but rapidly ageing real-time PCR infrastructure. Upgrading to state-of-the-art real-time PCR instrumentation in the Genomics Facility has become critical. Real-time PCR, which quantitatively detects increases in a PCR amplicon's copy number is a mainstay of modern molecular biology methods and supports a wide range of applications including functional genomics and gene expression, SNP detection, and species detection. It goes without saying that access to contemporary real-time PCR instrumentation is crucial to the continued success of a large number of research programs at UofG, from basic research in biodiversity science and human biology to applied advances in pest control and animal health and breeding. The existing StepOnePlus system in the Genomics Facility was purchased in 2004 and has been heavily utilized by the applicants and the facility's 100+ unique PIs and their 200+ students. The SetpOnePlus is approaching the end of its useful life. Users are experiencing frequent bottlenecks due to recurring downtime for repairs/maintenance due to instrument age. Critically, it also has limited flexibility in use that is impacting the competitiveness of research programs (e.g., limited to only a 96-well plate format). Because the instrument is increasingly down for repairs, it is highly oversubscribed when it is functional. The requested QuantStudio 7 Pro system offers maximum flexibility through an interchangeable thermal block system allowing for the use of 384-well plates and for the use of TaqMan Array microfluidics Cards (TAC). The system also allows for up to 6-dye multiplexing capability, which is the deepest on the market. The Orbitor RS2 microplate mover upgrade in addition to the QuantStudio's online run monitoring and data access will allow for 24/7 operation easing usage bottlenecks while simplifying user profile management. Setting up a high-end automated system like this in a core facility will help address the growing problem of sequestering and subsequent duplication of instrumentation at UofG while simultaneously supporting the research of the applicants and hundreds of other UofG researchers. The three core lab staff in the Genomics Facility provide real-time PCR training to users which consists of an introduction to real-time PCR, an explanation of lab workflows and relevant protocols, hands-on training on the operation of the QuantStudio and data analysis and interpretation.

高级分析中心（AAC）基因组学设施为整个UofG校园提供核心分子生物学技术支持服务和培训，其主要用户正在申请资助QuantStudio 7 Pro实时PCR系统，以取代现有但迅速老化的实时PCR基础设施。基因组学设施升级到最先进的实时PCR仪器已变得至关重要。实时PCR定量检测扩增子拷贝数的增加，是现代分子生物学方法的支柱，支持广泛的应用，包括功能基因组学和基因表达，SNP检测和物种检测。从生物多样性科学和人类生物学的基础研究到害虫防治和动物健康与育种的应用进展，获得现代实时PCR仪器对UofG大量研究项目的持续成功至关重要。基因组学设施现有的StepOnePlus系统于2004年购买，已被申请人和该设施的100多名独特的pi及其200多名学生大量使用。SetpOnePlus的使用寿命即将结束。由于仪器老化，用户经常遇到停机维修/维护的瓶颈。关键是，它在使用上的灵活性有限，影响了研究项目的竞争力（例如，仅限于96孔板格式）。由于越来越多的仪器需要维修，当它还能正常工作时，就会被大量超额认购。QuantStudio 7 Pro系统通过可互换的热块系统提供了最大的灵活性，允许使用384孔板和TaqMan阵列微流体卡（TAC）。该系统还允许多达6种染料的多路复用能力，这是市场上最深的。除了QuantStudio的在线运行监控和数据访问之外，Orbitor RS2微孔板移动器升级将允许24/7运行，缓解使用瓶颈，同时简化用户配置文件管理。在核心设施中建立这样一个高端自动化系统，将有助于解决UofG日益严重的仪器隔离和后续重复问题，同时支持申请人和数百名其他UofG研究人员的研究。基因组学设施的三名核心实验室工作人员为用户提供实时PCR培训，包括实时PCR的介绍，实验室工作流程和相关协议的解释，QuantStudio操作和数据分析和解释的实践培训。