A Quantitative platform to expedite gene and genomic discovery from model species to crops and wild berries.

定量、灵敏地检测基因组中基因表达和突变的变化对植物分子生物学研究至关重要。我们请求资助建立一个由定量实时PCR （qPCR）模块和集成数字PCR （dPCR）模块组成的平台，该平台将在不列颠哥伦比亚大学植物系托管。该qPCR模块是为了紧急替换一个失败的qPCR系统，该系统在过去十年中被UBC植物系大约一半的研究小组频繁使用。dPCR模块将支持qPCR无法解决的定量分析的相同用户组。dPCR的敏感性、精确性、抑制弹性和绝对定量允许检测罕见转录本、基因表达的微妙但生物学上至关重要的变化，以及自然发生的或基因组编辑诱导的突变，甚至在多倍体植物物种或PCR抑制剂存在的情况下。该定量平台将促进科学发现，并在至少8个NSERC发现资助的广泛研究项目中为50多个HQP的用户群体提供培训。这些研究活动将有利于植物分子生物学、细胞生物学、生物化学、代谢、基因组学和免疫等领域的许多基础研究项目。qPCR-dPCR平台还将产生定量数据，用于对加拿大具有高经济价值的物种的作物改良，包括油籽、小麦、蓝莓和西红柿，以及原住民利用的野生浆果。

Quantitative and sensitive detection of changes in gene expression and mutations in genomes is essential for plant molecular biology studies. We request funding to establish a platform composed of a quantitative real-time PCR (qPCR) module and an integrated digital PCR (dPCR) module to be hosted in the Department of Botany at the University of British Columbia. The qPCR module is for an urgent replacement of a failed qPCR system that was frequently used over the past decade by approximately half of the research groups in the Department of Botany at UBC. The dPCR module will support the same user groups for quantitative assays that cannot be addressed by qPCR. The sensitivity, precision, inhibition resilience, and absolute quantification of dPCR allow the detection of rare transcripts, subtle but biologically crucial changes of gene expression, and naturally occurring or genome editing-induced mutations even in polyploid plant species or in the presence of PCR inhibitors. The quantitative platform will facilitate scientific discoveries and training of more than 50 HQP across the user groups in a broad range of research programs supported by at least eight NSERC Discovery grants. These research activities will benefit the many fundamental research programs in plant molecular biology, cell biology, biochemistry, metabolism, genomics, and immunity. The qPCR-dPCR platform will also generate quantitative data for crop improvement in species of high economic value to Canada, including oilseeds, wheat, blueberries, and tomatoes, as well as wild berries utilized by First Nations.