This video documents large volume (≥20 L) filtration of microbial biomass, ranging between 0.22μm and 2.7μm in diameter, from the water column.
Representative Results: The final result consists of two Sterivix filters each containing microbial biomass from approximately 10 L seawater in the size range of 0.22µm and 2.7µm. This is stored in lysis buffer and ready for DNA extraction. In addition, this procedure collects and concentrates biomass greater than 2.7 µm, on the GF/D prefilters. This is also stored in lysis buffer and ready for DNA extraction. If additional processing steps are taken, the 0.22 µM flow-through can be concentrated down to collect the viral fraction as well.
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.
Material Name | Type | Company | Catalogue Number | Comment |
---|---|---|---|---|
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 Palmer | 0683204 | For filtration apparatus | |
1/4” x 3/16” pipe adaptors | Cole Palmer | 31702-20 | For filtration apparatus | |
Polypropylene prefilter housing | ADVANTEC | 501200 | For filtration apparatus | |
Metal luer fitting (Hose end to Male luer lock) | Cole Palmer | 31507-27 | For filtration apparatus | |
2“ Magnets | For filtration apparatus | |||
3 port carboy caps | For filtration apparatus | |||
GV 0.22 μm STERIVIX filters | Millipore | SVGV01R5 | ||
47 mm diameter 2.7 μm GF/D filters | Whatman | 1823047 | ||
Lysis buffer | Reagent | 0.75 M sucrose, 40 mM EDTA, 50 mM Tris (pH 8.3) | ||
0.1 % HCl | Reagent | For post-filtration rinse | ||
Autoclaved water | Reagent | For post-filtration rinse |