Team:INSA-Lyon/Protocols/Biofilms quantification
From 2010.igem.org
Protocols
Choose a protocol to read its description :
- Competent cells
- Transformation
- DNA extraction
- Digestion
- Ligation
- Measure of temperature and shaking speed influence
- Measure of osmotic pressure influence
- Granules extraction and intein cleavage
- Biofilms quantification
- Extra
Biofilms quantification
- Precultures of PHL818 and MG1655 in M63 medium + 0,2g/L glucose
- Dilute the cultures in order to have an absorbance (600nm) of 0,02, into a 24-well hydrophobic polystyrene plate, in (M63 + glucose)/2 medium, with sucrose concentrations varying between 0 and 0,3mol/L
- Incubate at 30°C, during 48h
- Pull out the 2ml supernatants and rinse one time with 1ml of M63 medium.
- Resuspend the adherent bacteria with 1ml of M63 medium.
- Measure the biomass in 3ml supernatants and also in 1ml biofilm (absorbance at 600nm). Calculate the percentage of bacteria in biofilms, which represents the adherence percentage.