Team:Imperial College/data
From 2013.igem.org
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<h2>Media characterisation</h2> | <h2>Media characterisation</h2> | ||
[[File:LB_M9.png]] | [[File:LB_M9.png]] | ||
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+ | {| class="wikitable" style="margin: 1em auto 1em auto;" | ||
+ | [[File:Waste_cocktail.png|thumbnail|center|600px|<b>(A)</b> WCM precursor material, this sterilised media made from LB and SRF was used to produce all WCM utilised. <b>(B)</b> Cells containing mCherry pigment grown in SRF <b>(A)</b> over 3 days, then streaked in a qualitative assay to check for growth. <b>(C)</b> mCherry cells were streaked again after 7 days growth in SRF.]] | ||
+ | [[File:PBS_Plus_Waste.jpg|thumbnail|center|600px|<b>(A)</b> SRF in PBS (phosphate buffered saline), a buffer. We can see from this experiment whether our bacteria can grow solely on the waste SRF. <b>(B)</b> Cells containing mCherry pigment grown in SRF <b>(A)</b> over 3 days, then streaked in a qualitative assay to check for growth. <b>(C)</b> mCherry cells were streaked again after 6 days growth in SRF.]] | ||
+ | |} | ||
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+ | <h2>pBAD characterisation</h2> | ||
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+ | {| class="wikitable" style="margin: 1em auto 1em auto;" | ||
+ | | [[File:Arabinose_induction.png|thumbnail|right|400px|Cell growth over 6h with IPTG induction. mCherry production is induced by the stress pathway and detection of ppGpps. In order to bypass this, we induced with IPTG which inhibits LacI, resulting in mCherry expression.]] | ||
+ | | [[File:F_arabinose_induction.png|thumbnail|right|400px|Fluorescence of the cells under IPTG induction over a 6h period.]] | ||
+ | |} | ||
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+ | {| class="wikitable" style="margin: 1em auto 1em auto;" | ||
+ | | [[File:Glucose_inhibition.png|thumbnail|right|400px|Cell growth over 6h with IPTG induction. mCherry production is induced by the stress pathway and detection of ppGpps. In order to bypass this, we induced with IPTG which inhibits LacI, resulting in mCherry expression.]] | ||
+ | | [[File:F_glucose_inhibition.png|thumbnail|right|400px|Fluorescence of the cells under IPTG induction over a 6h period.]] | ||
+ | |} | ||
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<h2>Glucose </h2> | <h2>Glucose </h2> | ||
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<br> | <br> | ||
- | < | + | <h3>WCM growth and IPTG induction assay</h3> |
Originally we intended on using [http://parts.igem.org/Part:BBa_K639003 BBa_K639003] to detect whether our cells were stressed when placed in various toxic byproducts. However, as the data below shows, this biobrick is very leaky. As such, we are using the stress sensor as a marker for cell growth and also to show that the cells had been successfully transformed with the correct chloramphenicol resistance. | Originally we intended on using [http://parts.igem.org/Part:BBa_K639003 BBa_K639003] to detect whether our cells were stressed when placed in various toxic byproducts. However, as the data below shows, this biobrick is very leaky. As such, we are using the stress sensor as a marker for cell growth and also to show that the cells had been successfully transformed with the correct chloramphenicol resistance. | ||
{| class="wikitable" style="margin: 1em auto 1em auto;" | {| class="wikitable" style="margin: 1em auto 1em auto;" | ||
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- | + | <h2>Sole carbon source</h2> | |
+ | <h3>3HB</h3> | ||
[[File:3HB_sole_carbon_source.png]] | [[File:3HB_sole_carbon_source.png]] | ||
- | + | <h3>Acetoacetate</h3> | |
[[File:AA_sole_carbon_source.png]] | [[File:AA_sole_carbon_source.png]] | ||
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[[File:M9M_phaCAB.png]] | [[File:M9M_phaCAB.png]] | ||
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<h1>PHB production</h1> | <h1>PHB production</h1> | ||
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Revision as of 00:06, 22 September 2013
Contents |
BioBricks
<groupparts>iGEM013 Imperial_College</groupparts>
Growth Assays
Media characterisation
pBAD characterisation
Glucose
Product inhibition on growth
L-lactic Acid
Ethylene glycol
3-hydroxybutyrate (3HB)
Acetoacetate
Poly(3-hydroxybutyrate) P(3HB)
Poly(lactic acid) (PLA)
WCM growth and IPTG induction assay
Originally we intended on using [http://parts.igem.org/Part:BBa_K639003 BBa_K639003] to detect whether our cells were stressed when placed in various toxic byproducts. However, as the data below shows, this biobrick is very leaky. As such, we are using the stress sensor as a marker for cell growth and also to show that the cells had been successfully transformed with the correct chloramphenicol resistance.
Sole carbon source
3HB
Acetoacetate
Induction
Empty Vector Control
Western blots
Enzyme Kinetics
PHB production