Disrupting the biofilm: Difference between revisions
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(New page: 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 compo...) |
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== Biofilm disruption solution == | == Biofilm disruption solution == | ||
* 1% SDS | * 1% SDS [http://en.wikipedia.org/wiki/Sodium_dodecyl_sulfate Sodium dodecyl sulfate] (denaturates protein) | ||
* 5 mM EDTA | * 5 mM [http://en.wikipedia.org/wiki/Edta EDTA] | ||
* 100 mM NaCl | * 100 mM NaCl | ||
* 50 mM Tris | * 50 mM [http://en.wikipedia.org/wiki/Tris Tris], pH 7.4, [[Tris | how to prepare]] | ||
== 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. |
Latest revision as of 14:29, 15 October 2007
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
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.