Difference between revisions of "Team:Calgary/BetaOxidation"

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<p><center><img src="https://static.igem.org/mediawiki/2017/5/57/Calgary2017_BetaOxidationPathway.png" alt="Beta-Oxidation Pathway" style="width:100%"></center></p>
 
<p><center><img src="https://static.igem.org/mediawiki/2017/5/57/Calgary2017_BetaOxidationPathway.png" alt="Beta-Oxidation Pathway" style="width:100%"></center></p>
  
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<h2>Aim</h2>
 
<h2>Aim</h2>
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<h2>Volatile fatty acids & beta-oxidation pathway</h2>
 
<h2>Volatile fatty acids & beta-oxidation pathway</h2>
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<h2>Gene construct</h2>
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<h2>Genetic construct</h2>
  
  
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<h2> Experiment </h2>
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<h2>Results</h2>
 
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 OD<sub>600</sub>. The composition of each of the replicate in the flasks is shown below:</p>
 
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 OD<sub>600</sub>. The composition of each of the replicate in the flasks is shown below:</p>
 
<table>
 
<table>

Revision as of 05:46, 31 October 2017

Header

Beta Oxidation

Beta-Oxidation Pathway


Aim

...bla bla bla


Volatile fatty acids & beta-oxidation pathway

…


Genetic construct

Beta-Oxidation Construct

…


Results

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:

Glucose (Positive control) pET29B in BL21 (Negative control)
  • 10 ml O/Ns in LB+Kan
  • 7 ml 20% Glucose
  • 100 uL MgSO4
  • 5 uL CaCl2
  • 10 ml M9 salts
  • 5 uL 1M IPTG
  • 20 ml dH2O
  • 10 ml O/Ns in LB+Kan
  • "Syn poo" fermented supernatant
  • 100 uL MgSO4
  • 5 uL CaCl2
  • 10 ml M9 salts
  • 5 uL 1M IPTG
Fermented "syn poo" supernatant Pure VFAs
  • 10 ml O/Ns in LB+Kan
  • 10 ml "Syn poo" fermented supernatant
  • 100 uL MgSO4
  • 5 uL CaCl2
  • 10 ml M9 salts
  • 5 uL 1M IPTG
  • 23 ml dH2O
  • 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
  • 5 uL 1M IPTG
  • 30 ml dH2O

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