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

对基因表达变化和基因组突变的定量和灵敏检测是植物分子生物学研究的基础。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省大学植物系托管。qPCR模块用于紧急更换失败的qPCR系统，该系统在过去十年中经常被UBC植物系大约一半的研究小组使用。dPCR模块将支持qPCR无法解决的定量检测的相同用户组。dPCR的灵敏度、精确度、抑制恢复力和绝对定量允许检测罕见的转录本、基因表达的微妙但生物学上至关重要的变化以及甚至在多倍体植物物种中或在PCR抑制剂存在下天然发生的或基因组编辑诱导的突变。该定量平台将促进科学发现和培训超过50个HQP的用户群体在广泛的研究计划支持至少八个NSERC发现赠款。这些研究活动将有益于植物分子生物学、细胞生物学、生物化学、代谢、基因组学和免疫学的许多基础研究计划。qPCR-dPCR平台还将生成对加拿大具有高经济价值的作物改良的定量数据，包括油籽、小麦、蓝莓和番茄，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.