Colón-Padilla, Brenda L.

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    Development of an autonomous sample filtration and archival device for aquatic microbiology: Assessment of potential preservation buffers
    (2011) Colón-Padilla, Brenda L.; Smith, Matthew; College of Arts and Sciences - Sciences; Schizas, Nikolaos V.; Otero, Ernesto; Aponte, Nilda; Department of Marine Sciences; Gilbes Santaella, Fernando
    Ongoing advances in molecular techniques have certainly increased our understanding of marine microbial ecology. However, much of this knowledge could be enhanced by increased sampling frequency. This work presents the first development phase of a small autonomous in situ water sampling device. The system will enable the autonomous collection of water samples to be preserved for subsequent downstream laboratory based molecular techniques. The Sample Filtration and Archiving (SaFA) system is being designed to collect and filter 24 user defined time-stamped water samples of between ~200-500 ml. During this phase, simulated SaFA tests using several commercial and noncommercial buffers were conducted to evaluate their capacity to preserve and stabilize DNA from natural seawater samples. The best performers of the commercially available buffers were PrepProtect, RNALater, Ethanol, and Isopropanol. These buffers were effective for 3 to 8 days without appreciable DNA degradation and significant microbial community changes. Only one non-commercial buffer (HRNAL pH 5.2) was efficient up to 5 days however this formulation represents a homemade version of the commercially available RNALater. These results indicate that future deployment of SaFA could be limited for periods up to a week under the condition of our tests. To facilitate adoption by end users, the SaFA is being constructed using predominately commercially available parts, and all information relating to its construction and operation will be downloadable under an open source license. The open source nature of the SaFA will enable end users to evolve the instrument by providing feedback on new designs or applications.