DNA isolation from dislodged biofilms was performed as described previously [41 (link)]. Briefly, biofilm pellets were thawed, resuspended with 150 μl Tris EDTA buffer (10 mM Tris-Cl (pH 8.0), 1 mM EDTA (pH 8.0)) and transferred to a well in a 96 deep well plate (Axygen Scientific Inc., CA, USA). As controls, the original inoculum and sterile McBain medium without additions from each experiment were included.
To each well, the following compounds were added: 250 μl of 0.1-mm diameter Zirconia beads (BioSpec Products, Bartlesville, OK, USA), 200 μl of phenol (Rotiphenol, Carl Roth GMBH&Co. KG, Germany) and 200 μl of lysis buffer (MagMini DNA isolation kit, LGC Genomics Ltd, UK). The plate was then sealed and placed in a Mini-BeadBeater-96 (BioSpec Products, Bartlesville, OK, USA) for 2 min at 2.100 oscillations/min. When this process was completed, DNA was extracted and purified with the MagMini DNA Isolation Kit (MagMini DNA isolation kit, LGC Genomics Ltd, UK). Bacterial DNA concentration was determined by qPCR as described elsewhere [42 (link)].
To each well, the following compounds were added: 250 μl of 0.1-mm diameter Zirconia beads (BioSpec Products, Bartlesville, OK, USA), 200 μl of phenol (Rotiphenol, Carl Roth GMBH&Co. KG, Germany) and 200 μl of lysis buffer (MagMini DNA isolation kit, LGC Genomics Ltd, UK). The plate was then sealed and placed in a Mini-BeadBeater-96 (BioSpec Products, Bartlesville, OK, USA) for 2 min at 2.100 oscillations/min. When this process was completed, DNA was extracted and purified with the MagMini DNA Isolation Kit (MagMini DNA isolation kit, LGC Genomics Ltd, UK). Bacterial DNA concentration was determined by qPCR as described elsewhere [42 (link)].
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