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 (denaturates protein)
- 5 mM EDTA
- 100 mM NaCl
- 50 mM Tris, pH 7.4, how to prepare
- 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.