Team:Groningen/13 September 2010

From 2010.igem.org

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Chaplinj were added as a extra control.
Chaplinj were added as a extra control.
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'''Exression experiment'''
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'''Peter & David'''
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For this experiment, the following B. subtilis 168 strains were used:
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<pre>
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</pre>
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All cultures were grown overnight at 37 degrees Celsius in a shaker room, the appropriate antibiotics were used at all points in time during this experiment.
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Overnight cultures were  used to dilute to a B. subtilis culture of 0,1 OD, these strains were divided into ‘’induced’’ and ‘’non-induced’’. Induction with 0,5% subtilin was done at a OD of 0,5 (approximately 2,5 hours after growth of the 0,1 culture started).
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After that the OD of the cultures was measured every .. hours.
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'''Sample preperation'''
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After .. hours, .. after induction, the samples were collected and processed. The following procedures were used:
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<br>
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Pellet preperation ([[PelletPrepGR]])
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Supernatant processing ([[SupernatantPrepGR]])
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Cell disruption ([[ExtractionCellWallsGR]])
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Lysozyme preperation ([[LysozymePrepGR]])
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Analysis was done using SDS-PAGE ([[SDS-PAGEGR]]) and THT staining ([[THTstainingGR]]).
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<br>
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'''Results''':
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'''Growth Curve'''
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[[Image:GrowthCurveGR3.jpg|400px]]
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'''THT Staining'''
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[[Image:THTGR3.jpg|400px]]
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'''SDS-PAGE'''
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[[Image:SDS-PAGEGR3.jpg|400px]]
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{{Team:Groningen/Footer}}
{{Team:Groningen/Footer}}

Revision as of 16:14, 24 October 2010

iGEM Groningen 2010

Hydrophobofilm
pushing coatings into a greener future


Week 27
Expression experiment - David & Peter
A broad experiment was set up: lysis/disrupted/differentconcentrationsubtilin/delta.
No positive results were found, a small sample size.
A experiment using anti-biotics as well, growth curves were interesting.
Chaplinj were added as a extra control.

Exression experiment


Peter & David


For this experiment, the following B. subtilis 168 strains were used:


All cultures were grown overnight at 37 degrees Celsius in a shaker room, the appropriate antibiotics were used at all points in time during this experiment.


Overnight cultures were used to dilute to a B. subtilis culture of 0,1 OD, these strains were divided into ‘’induced’’ and ‘’non-induced’’. Induction with 0,5% subtilin was done at a OD of 0,5 (approximately 2,5 hours after growth of the 0,1 culture started).


After that the OD of the cultures was measured every .. hours.


Sample preperation


After .. hours, .. after induction, the samples were collected and processed. The following procedures were used:


Pellet preperation (PelletPrepGR)


Supernatant processing (SupernatantPrepGR)


Cell disruption (ExtractionCellWallsGR)


Lysozyme preperation (LysozymePrepGR)


Analysis was done using SDS-PAGE (SDS-PAGEGR) and THT staining (THTstainingGR).


Results:


Growth Curve


GrowthCurveGR3.jpg




THT Staining


400px




SDS-PAGE


400px




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