Team:Groningen/20 September 2010

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iGEM Groningen 2010

Hydrophobofilm
pushing coatings into a greener future

Week 29

Expression test-Peter & David

Succesfull in disrupted samples!

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


GrowthCurveGR6.jpg




THT Staining


Error creating thumbnail: Image type not supported




SDS-PAGE


400px




Chaplin ladder on gel-Peter

Modellers: Find constants and parameters for gene expression model Laura, Djoke

Initial programming of gene expression model in Matlab. Laura



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