Replacement of automated real-time PCR cluster

更换自动化实时 PCR 集群

• 批准号: RTI-2021-00413
• 负责人: AbouElela, Sherif
• 金额: $ 10.47万
• 依托单位: Université de Sherbrooke
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=718838
• 关键词: Replacement; automated; real; time; PCR

The capacity to amplify nucleic acid is an essential process of life sciences and the backbone of molecular biology research. The ability to quickly and precisely amplify from RNA or DNA several gene markers in high throughput provides powerful data for understanding the mechanism of gene expression and determining the regulatory elements that affect RNA production and stability. By using set of real-time thermocyclers, we developed methods for high-throughput analysis of the different RNA biotypes including hard to detect transcripts like tRNA, miRNA, snoRNA and splice variants. Using an integrated nucleic acid detection and analysis pipeline we have developed optimized methods for transcriptome wide quantification of gene expression, validation of NextGen sequencing data, sequencing library quantification, identification and classification of microorganism (e.g. bacteria, viruses and fungi), mycoplasma detection, quantification of splice variants, detection of cell culture and animal infections, microbiome analysis, telomerase assay and the detection of repetitive and duplicated genes. These newly developed methods are now being used by many researchers worldwide. Indeed, our real-time PCR rapidly became the gateway of the Université de Sherbrooke RNomics platform (https://rnomics.med.usherbrooke.ca/) that caters to the needs of basic researchers interested in RNA biology and genomics. The goal of this application is to acquire replacements for the nucleic acid real-time PCR equipment needed to maintain and upgrade the service of the RNomics platform. It is essential for our institution to maintain its capacity to provide high-throughput real-time PCR in order to prevent the disruption of the NSERC-funded research programs requiring the detection and measure of nucleic acid levels. We are requesting funds to replace the six real-time PCR machines that we have used for the last 12 years to support the study of RNA expression. The existing machines stopped working are discontinued and no spare-parts are available for repair. There are no similar platform set-up available on campus and without it the whole pipeline of nucleic-acid analysis required to complete our NSERC-funded research will cease to function. The new machines will ensure continued service of at least 35 labs at the Université de Sherbrooke and over 100 different researchers who are using the RNomics platform nationally and internationally. Most importantly, the requested equipment will permit the continuation of our unique graduate training programs focused on RNA biology. In short, the requested machines are absolutely needed as replacements for basic equipment essential for supporting the research and training program funded by NSERC.

核酸扩增能力是生命科学的基本过程，也是分子生物学研究的支柱。快速、准确、高通量地从RNA或DNA中扩增多个基因标记的能力为理解基因表达的机制和确定影响RNA生产和稳定性的调控元件提供了强大的数据。通过使用一套实时热循环仪，我们开发了高通量分析不同RNA生物型的方法，包括难以检测到的转录本，如tRNA、miRNA、snoRNA和剪接变体。使用集成的核酸检测和分析流水线，我们开发了优化的方法，用于基因表达的转录组范围的量化、NextGen测序数据的验证、测序文库的量化、微生物(例如细菌、病毒和真菌)的鉴定和分类、支原体检测、剪接变异体的定量、细胞培养和动物感染的检测、微生物组分析、端粒酶分析以及重复和重复基因的检测。这些新开发的方法现在正被世界各地的许多研究人员使用。事实上，我们的实时聚合酶链式反应迅速成为舍布鲁克大学核糖核酸组学平台(https://rnomics.med.usherbrooke.ca/))的门户，满足了对核糖核酸生物学和基因组学感兴趣的基础研究人员的需求。这项应用的目标是获得维护和升级RNome平台服务所需的核酸实时聚合酶链式反应设备的替代品。对于我们的机构来说，保持其提供高通量实时聚合酶链式反应的能力是至关重要的，以防止NSERC资助的要求检测和测量核酸水平的研究计划的中断。 我们正在申请资金，以更换我们在过去12年中一直使用的六台实时PCR机，以支持RNA表达的研究。已停止工作的现有机器已停产，没有备件可供维修。校园里没有类似的平台，没有它，完成我们NSERC资助的研究所需的整个核酸分析管道将停止运转。这些新机器将确保舍布鲁克大学的至少35个实验室和100多名不同的研究人员继续使用RNome平台，这些研究人员将在国内外使用该平台。最重要的是，所要求的设备将允许我们继续开展以RNA生物学为重点的独特的研究生培训计划。简而言之，所要求的机器是绝对需要的，作为支持NSERC资助的研究和培训计划所必需的基本设备的替代品。