Team:INSA-Lyon/Protocols/Measure of osmotic pressure influence

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Protocols


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  1. Competent cells
  2. Transformation
  3. DNA extraction
  4. Digestion
  5. Ligation
  6. Measure of temperature and shaking speed influence
  7. Measure of osmotic pressure influence
  8. Granules extraction and intein cleavage
  9. Biofilms quantification
  10. Extra






    Measure of osmotic pressure influence


      Preparation of different media : 0M - 0,01M - 0,05M - 0,1M – 0,5M
      1. x (µL) Sucrose 1M
      2. 2,5mL media M63
      3. 50µL of glucose
      4. Complet with steril water until 5mL

      Incubate during 24 h at the temperature 28°C and the shaking speed of 100rpm

      Measure of the fluorescence : input wavelenght 490 nm, output one 510 nm.

      Measure of the biomass(absorbance 600 nm): Dilute 250 µL of culture in 750 µL of water.
      Make the blank with 0,5mL of media M63 and 0,5mL of steril water

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