Department of Microbiology and Immunology, University of British Columbia - UBC
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Walsh, D. A., Zaikova, E., Hallam, S. J. Small Volume (1-3L) Filtration of Coastal Seawater Samples. J. Vis. Exp. (28), e1163, doi:10.3791/1163 (2009).
Representative Results: Each 1 L water sample yields one Sterivix filter, which contains biomass greater than 0.22 µm. This is stored in lysis buffer for subsequent DNA extraction and analysis. The final result consists of one Sterivix filter per 1-liter bottle stored in lysis buffer and ready for DNA extraction. The filter contains biomass greater than 0.22 µm. Filters can be stored at -80°C prior to eDNA extraction.
It is imperative to wipe down benches and pumps with a damp towel after use to remove any lingering traces of salt water that would otherwise corrode equipment. Although the procedure is simple, the time requirement can be extensive depending on the concentration of biomass or debris in the sample being filtered.
The authors have nothing to disclose.
We would like to thank the Canadian Foundation for Innovation, the British Columbia Knowledge Development Fund and the National Sciences and Engineering Research Council (NSERC) of Canada for supporting ongoing studies on low oxygen regions of coastal and open ocean waters. D.A.W. was supported by fellowships from NSERC, Killam and the TULA foundation funded Centre for Microbial Diversity and Evolution.
|Peristaltic pump||Masterflex (Cole Palmer)||77913-20, 77200-62, 77200-04|
|Tygon LFL tubing||Masterflex (Cole Palmer)||06429-24||For filtration apparatus|
|Hose clamps||Cole-Parmer||0683204||For filtration apparatus|
|1/4” x 3/16” pipe adaptors||Cole-Parmer||31702-20||For filtration apparatus|
|Polypropylene prefilter housing||ADVANTEC||501200||For filtration apparatus|
|Metal luer fitting (Hose end to Male luer lock)||Cole-Parmer||31507-27||For filtration apparatus|
|2“ Magnets||For filtration apparatus|
|3 port carboy caps||For filtration apparatus|
|GV 0.22 μm STERIVIX filters||EMD Millipore||SVGV01R5|
|47 mm diameter 2.7 μm GF/D filters||Whatman, GE Healthcare||1823047|
|Lysis buffer||0.75 M sucrose, 40 mM EDTA, 50 mM Tris (pH 8.3)|
|0.1 % HCl||For post-filtration rinse|
|Autoclaved water||For post-filtration rinse|