Replacement of automated real-time PCR cluster

点击翻译按钮获取中文摘要

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.