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JOHNS HOPKINS UNIVERSITY, THE

The funds requested will be used to purchase a Tecan Infinite M200 Fluorescence plate reader with shaking platform and temperature control. This highly versatile instrument will enhance our abilities to fulfill all three aims of our current NIH R01 grant GM60979 entitled Molecular Analysis of Endocytosis Proteins in Yeast. For Aims 1 and 2, we have proposed to conduct yeast two-hybrid assays, protein binding assays, and analyses of endocytosis mutant strains of yeast cells. For each of these experimental approaches, using the Tecan M200 will allow us to do high-throughput, quantitative assays in a much more efficient and precise manner. For Aim 3, we have proposed to do protein binding assays that again will benefit from using the Tecan M200 for more quantitative readouts. Our lab arranged for a demo unit, which proved to us that this instrument will be of great benefit to our experimental productivity. Further, as is our usual approach, we will be happy to share access of this instrument with other NIH funded investigators in our department, thus furthering the value received by the NIH for providing funds to allow us to purchase this instrument. We are also requesting funds to purchase some minor replacement equipment for basic operation of our laboratory. As our lab has been in operation for eleven years, the usual wear-and-tear on some basic equipment is beginning to take its toll. We have already replaced numerous items over the past two years, and some assistance with replacing an ultracentrifuge rotor for our benchtop ultracentrifuge (used almost daily for Aim 3 of our NIH R01 grant), and replacement of some of our worn out variable volume pipetters (used many times per day for all three Aims), would be of great help.

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