Disrupting the biofilm

The crystal violet assay is great to quantify biofilm formation quickly in a high-throughput way for single- and multiple-strain biofilms. Nevertheless, we want to look at the strain composition in each biofilm in a high. For this, we need to disrupt the biofilm back into suspended cells so that we can quantify their relative amounts through fluorescence.

Biofilm disruption solution

• 1% SDS Sodium dodecyl sulfate
• 5 mM EDTA
• 100 mM NaCl
• 50 mM Tris
• pH 7.4

Protocol

• Grow biofilm in 96-well plate.
• Extract liquid (measure OD if necessary) and wash twice.
• Fill with 150 μm disruption solution.
• Shake for 30 mins.
• Measure OD.
• Measure fluorescences.