Team:Calgary/BetaOxidation

Header

Beta Oxidation

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Aim

...bla bla bla

Volatile fatty acids & beta-oxidation pathway

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Gene construct

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Chassis and vector

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Experiment

The O/Ns were grown in the respective media for ~24 hours. The flasks containing different media was inoculated with the O/Ns after adjusting the OD600. The composition of each of the replicate in the flasks is shown below:
pET29B in BL21 (Negative control)
    10 ml O/Ns in LB+Kan
    "Syn poo" fermented supernatant {..!!hyperlink to eng page for syn poo protocol..}
    100 uL MgSO4
    5 uL CaCl2
    10 ml M9 salts
Glucose (Positive control)
    10 ml O/Ns in LB+Kan
    7 ml 20% Glucose {..!!cite why this conc chosen..}
    100 uL MgSO4
    5 uL CaCl2
    10 ml M9 salts
Fermented "syn poo" supernatant
    10 ml O/Ns in LB+Kan
    10 ml "Syn poo" fermented supernatant {..!!hyperlink to eng page for syn poo protocol..}
    100 uL MgSO4
    5 uL CaCl2
    10 ml M9 salts
Pure VFAs
    10 ml O/Ns in LB+Kan
    VFAs
    410 uL propionic acid
    118 uL acetic acid
    55 uL butyric acid
    100 uL MgSO4
    5 uL CaCl2
    10 ml M9 salts

The OD600 readings of .....???? were taken and recorded:

Condition OD600 of replicate 1 OD600 of replicate 2 OD600 of replicate 3
pET29b in BL21 (Negative control) 0.571 0.531 0.487
Glucose (Positive control) 0.190 0.195 0.139
Pure VFAs 0.140 0.134 0.146
Fermented "syn poo" supernatant 0.135 0.107 0.144

After spinning down the culture in flasks, the cells were resuspended in 1x PBS. The OD600 readings were taken:

Condition OD600 of replicate 1 OD600 of replicate 2 OD600 of replicate 3
pET29b in BL21 (Negative control) 2.659 2.001 2.899
Glucose (Positive control) 1.934 1.887 1.919
Pure VFAs 0.510 0.571 0.532
Fermented "syn poo" supernatant 2.533 2.559 2.349