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Team:Groningen/20 September 2010
From 2010.igem.org
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
When preparing the pellets for disruption there was a difference visible between the induced and the non induced strains. The induced sample on the left formed a somewhat flaky pellet in comparison to the non induced sample.
THT Staining
SDS-PAGE
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
