Real-time quantitative PCR instrument

本提案要求资金购买定量实时PCR（聚合酶链反应）的分析系统。这种类型的仪器已成为分子生物学实验室基因表达研究的必要条件。由于其灵敏度，更准确地量化mRNA水平的能力，以及分析的速度和特异性，实时PCR通常被认为优于RNA (northern) blots等传统技术。该仪器将主要用于植物系几个实验室的基因表达分析，使用定量反转录PCR分析（qPCR）。基于实时的qPCR基因表达分析是我们实验室在植物细胞和发育生物学以及植物生物化学领域开展的许多研究的核心和关键分析类型。该仪器的具体应用将是获得与细胞壁生物量生产、植物蜡和油的生物合成、植物细胞骨架的细胞生物学以及海洋硅藻生产力研究等不同过程相关的关键基因的表达谱。虽然qPCR是必不可少的五个共同申请者，但我们的实验室目前都没有这样的仪器，我们依赖于位于我们部门另一个实验室的一个频繁使用的仪器。由于所有这些仪器的大量使用及其容量限制，我们在进行对我们的研究项目至关重要的基因表达研究时，在研究进展和生产力方面面临瓶颈和潜在的延迟。所要求的实时qPCR系统将缓解这些瓶颈，不仅为我们的四个实验室提供集中资源，而且根据需要为本部门的其他实验室提供资源。

This proposal requests funds to purchase an analytical system for quantitative real time PCR (polymerase chain reaction). Instruments of this type have become essential in molecular biology labs for gene expression studies. Real-time PCR is generally regarded as superior to traditional techniques such as RNA (northern) blots, due to its sensitivity, ability to more accurately quantify mRNA levels, and due to the speed and specificity of the analyses. The instrument will be mainly used for gene expression analysis using quantitative reverse transcription PCR assays (qPCR) in several labs in the Department of Botany.  Real-time based qPCR gene expression assays are central and key types of analyses fundamental to many aspects of the research carried out in our labs in the areas of plant cell and developmental biology and plant biochemistry. Specific applications of this instrument will be to obtain expression profiles of key genes associated with diverse processes in cell wall biomass production, biosynthesis of plant waxes and oils, the cell biology of the plant cytoskeleton, and studies on the productivity of marine diatoms. While qPCR is essential the five co-applicants, none of our labs currently has such an instrument, and we rely on a single heavily used instrument located in another lab in our department. Due to the heavy use of all of this instrument, and its capacity limitations, we face bottlenecks and potential delays in research progress and productivity in carrying out gene expression studies essential to our research programs. The requested real time qPCR system will alleviate these bottlenecks, and provide a centralized resource available not only to our four labs, but to others in the Department as needed.