Real-time quantitative PCR instrument

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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.