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

高级分析中心(AAC)基因组学设施为整个UofG园区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。在基因组学设施中升级到最先进的实时聚合酶链式反应仪器已经变得至关重要。实时定量聚合酶链式反应是现代分子生物学方法的主流，它可以定量检测聚合酶链式反应扩增产物的拷贝数，支持功能基因组学和基因表达、单核苷酸多态性检测和物种检测等广泛的应用。不用说，获得当代实时PCR仪器对于UofG的大量研究项目的持续成功至关重要，从生物多样性科学和人类生物学的基础研究到虫害控制、动物健康和育种的应用进展。基因组学设施中现有的StepOnePlus系统是在2004年购买的，申请者和该设施的100个独特的PI及其200名学生大量使用该系统。SetpOnePlus的使用寿命即将结束。由于仪器老化导致维修/维护的反复停机，用户经常遇到瓶颈。关键的是，它在使用上的灵活性也是有限的，这影响了研究计划的竞争力(例如，仅限于96孔板格式)。由于该仪器越来越需要维修，因此当它正常工作时，它的认购量会大幅超额。所要求的QuantStudio 7 Pro系统通过可互换的热块系统提供了最大的灵活性，该系统允许使用384孔板和TaqMan阵列微流体卡(TAC)。该系统还允许高达6染料的多路复用能力，这是市场上最深入的。Orbitor RS2微板移动器的升级，加上QuantStudio的在线运行监控和数据访问，将允许全天候运行，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的高端自动化系统将有助于解决UofG日益严重的隔离和随后的仪器重复问题，同时支持申请者和数百名其他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向用户提供实时聚合酶链式反应培训，包括对实时聚合酶链式反应的介绍，对实验室工作流程和相关协议的解释，关于QuantStudio操作以及数据分析和解释的动手培训。

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.