Team:Imperial College London/Wetlab

Assay Rationale:

Colanic acid biosynthesis is both time consuming and metabolically expensive. If the E.ncapsulator is to be used in an industrial setting, then colanic acid mediated protection must be highly efficient. Protective efficiency can be defined as the percentage increase in fluorescence per µl of colanic acid produced (when compared to control cells). The following assay can be used to elucidate this parameter.

Assay Overview:

1) Growth of cells (Assay preparation).

2) Measurement of cell density.

3) Measurement of packed cell volume (PCV).

4) Measurement of the % change in GFP fluorescence following acid incubation.

DAY 1:

AM:

M9 Minimal Media Preparation:

Measure out the following reagents and dissolve them in 1000ml of sterile H20:

Disodium Phosphate = 6.0g

Potassium dihydrogen phosphate = 3.0g

Sodium Chloride = 0.5g

Ammonium Chloride = 1.0g

Carbon source (as determined by Kun) = 4.0g

Casamino acids =?

Leave out for autoclaving (before 12:30)

PM:

Collect media from autoclave (at 5pm)

M9 Minimal Media Innoculation:

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