A Real-Time Solution for a Kinetic Challenge
Dr. Gautam Dantas' lab is part of Washington University School of Medicine's newly renovated Center for Genome Sciences and Systems Biology. The open-format and modular lab space is setup to enable interdisciplinary and collaborative research with neighboring labs. The Dantas lab seeks to understand genetic and biochemical mechanisms that allow some microbes to tolerate materials that are toxic to other microbes. This research may be used to understand and combat antibiotic resistance and to improve biofuel production.
Kinetic growth studies focus on toxin-tolerating bacteria and genetically modified E. coli (where a gene of interest from another bacteria is inserted) to assess phenotype expression under various conditions. With so many combinations to explore, these kinetic studies were a bottleneck to Dr. Dantas’ research. "We did end-point studies at my previous lab, but that doesn’t tell the whole story; it doesn’t address what happens during the growth cycle," notes Dr. Dantas. "To understand the entire growth process and get the most accurate data, we needed to run high-throughput kinetic assays." This application required reading each microplate well every 40 minutes, and was generally limited to a single 96-well microplate per day.
In the course of setting up his new lab, Dr. Dantas relayed his kinetic growth study needs to his local distributor who recommended a few instrument/software manufacturers, including BioTek Instruments. BioTek’s local sales representative, Jason Fries, met with Dr. Dantas, and the two worked with BioTek’s software engineers to create a Gen5™ Data Analysis Software protocol to run multiple kinetic assays and generate real-time growth curves using the PowerWave™ Microplate Spectrophotometer and the BioStack™ Microplate Stacker. A microplate sealing film is used instead of a lid to maintain gas exchange while preventing contamination.
The new system significantly improves efficiency, processing up to fifty 96-well plates per day compared to the old method’s throughput of a single plate per day. The plates are read every 40 minutes, with subsequent generation of up to 4800 real-time growth curves.
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