Replacement of automated real-time PCR cluster

扩增核酸的能力是生命科学必不可少的过程，也是分子生物学研究的支柱。从RNA或DNA中快速精确地高通量扩增多个基因标记的能力为理解基因表达机制和确定影响RNA产生和稳定性的调控元件提供了强大的数据。通过使用一组实时热循环仪，我们开发了高通量分析不同RNA生物型的方法，包括难以检测的转录本，如tRNA， miRNA， snoRNA和剪接变体。利用集成的核酸检测和分析管道，我们开发了优化的方法，用于基因表达的转录组定量、NextGen测序数据的验证、测序文库定量、微生物（如细菌、病毒和真菌）的鉴定和分类、支原体检测、剪接变异体定量、细胞培养和动物感染检测、微生物组分析、端粒酶测定及重复和重复基因的检测。这些新开发的方法目前正在被世界各地的许多研究人员使用。事实上，我们的实时PCR迅速成为Sherbrooke大学RNomics平台（https://rnomics.med.usherbrooke.ca/）的门户，该平台迎合了对RNA生物学和基因组学感兴趣的基础研究人员的需求。本申请的目的是获得维护和升级RNomics平台服务所需的核酸实时PCR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PCR的能力是至关重要的，以防止nserc资助的需要检测和测量核酸水平的研究项目中断。

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.