Exploring cyclic di-nucleotide signaling across the tree of life (Supplement)

I am requesting support for the purchase of an Agilent BioTek LogPhase 600 microbiology plate reader through Notice NOT-GM-24-021 under the parent grant R35 GM139537-01, “Exploring cyclic di-nucleotide signaling across the tree of life”. This equipment allows for simultaneous quantification of the optical density at 600 nm

(OD600) of bacteria in four 96-well plates. The LogPhase 600 will be used to measure bacterial growth curves, which is one of the most common and fundamental phenotypes assayed in a microbiology laboratory is measuring growth kinetics by tracking optical density, usually at a wavelength of 600 nm (OD600), over time. A

growth curve can yield the growth rate of bacteria, their lag time at the initiation of the culture, when they enter stationary phase and stop growing, and the maximum yield of the population. Such values are fundamental in understanding the overall physiology of bacterial cells. We also use such growth curves to study the interaction

of bacteria with their cognate viruses known as phage. When phage infects a bacterial population, it leads to lysis of the bacterial cells, which can be measured by a drop in the OD600. To survive the onslaught of phage that bacteria encounter in the natural world, they have evolved a myriad of molecular defense systems. Some of

these systems, like restriction enzymes and CRISPR, have been studied for decades and are now fundamental molecular tools for modifying and manipulating DNA. However, over the last ~10-years, a wide range of diverse defense systems has been described, including three from my lab, resulting in several publications since the

start of the R35. A major bottleneck for this research is access to a plate reader that can track OD600 over time in a high-throughput fashion. I chose the Agilent BioTek LogPhase 600 microbiology plate reader as it is a simple device that does one thing well-measure OD600 over time in 96-well plates. The most exciting part about this

piece of equipment is that it can analyze four plates simultaneously. This allows up to four members of my lab to perform growth curves or phage infection studies at the same time, or one member to do multiple replicates of one experiment. Having access to the LogPhase 600 would significantly advance the progress of research in

my lab, and at a cost of only $34,245.70, it is a relatively low-cost piece of equipment that will be give a large “bang for the buck”. The investment in this equipment by the NIH would greatly enhance the return on my R35 GM139537-01 award.