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 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金购买 QuantStudio 7 Pro 实时 PCR 系统，以取代现有但快速老化的实时 PCR 基础设施。基因组设施升级至最先进的实时 PCR 仪器已变得至关重要。实时 PCR 可定量检测 PCR 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。不用说，使用现代实时 PCR 仪器对于乔治大学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害虫控制、动物健康和育种的应用进展。 基因组学设施中现有的 StepOnePlus 系统于 2004 年购买，已被申请人、该设施的 100 名独特 PI 及其 200 名学生大量使用。 SetpOnePlus 的使用寿命即将结束。 由于仪器老化导致维修/维护反复停机，用户经常遇到瓶颈。重要的是，它的使用灵活性也有限，影响了研究项目的竞争力（例如仅限于 96 孔板格式）。由于该仪器因维修而停机的情况越来越多，因此当其正常运行时，其订购量会很高。 所要求的 QuantStudio 7 Pro 系统通过可互换的热块系统提供最大的灵活性，允许使用 384 孔板和 TaqMan 阵列微流体卡 (TAC)。该系统还支持多达 6 种染料的多重分析能力，这是市场上最深入的。除了 QuantStudio 的在线运行监控和数据访问之外，Orbitor RS2 微孔板移动器的升级将允许 24/7 操作，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乔治大学日益严重的仪器隔离和随后重复的问题，同时支持申请人和数百名其他乔治亚大学研究人员的研究。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 简介、实验室工作流程和相关协议的解释、QuantStudio 操作以及数据分析和解释的实践培训。

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.