An Automated, Pressure-Driven Sampling Device for Harvesting from Liquid Cultures for Genomic and Biochemical Analysis
Anthony D. Aragon1, Gabriel A. Quinones1, Chris Allen1, Jason Thomas1, Sushmita Roy2, George S. Davidson3, Peter D. Wentzell4, Brian Millier4, Jason E. Jaetao1, Angelina L. Rodriguez1, and Margaret Werner-Washburne1
1Department of Biology, University of New Mexico, Albuquerque, NM 87131, 2Department of Computer Science, University of New Mexico, Albuquerque, NM 87131, 3Sandia National Laboratories, Albuquerque, NM 87185, 4Department of Chemistry, Dalhousie University, Halifax, NS B3H 4J3, Canada. Correspondence should be addressed to M.W.W. (firstname.lastname@example.org).
Routine, sterile sampling of intact cells from liquid cultures, in volumes needed for biochemical and genomics analyses, and with time points ranging from seconds to minutes presents a significant challenge for systems biology. Here we describe a relatively simple, automated, pneumatic-sampling device for harvesting up to 30-ml of liquid, in replicate, with sampling intervals as short as 10 s. The simplicity and modularity of the sampler allows it to be coupled with a variety of harvesting methods, e.g. filtration, centrifugation, etc. This device was used to study an extremely rapid increase in transcript abundance, in yeast cells in stationary-phase cultures, in response to oxidative stress. Correlation between biological replicate time courses harvested with this device measured by microarrays was extremely high. The sampler reported here enables robust, real-time, integrated studies at time intervals within the range of many important biological processes.
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