Part:BBa_K1951010:Experience

Test of our biobrick proteins production using a SDS page and comassie

We investigated if the FliC protein was well produced by our biobrick using SDS PAGE.

To to do this we performed SDS PAGE and stained with coomassie blue using cells containing this biobrick in plasmid backbone [http://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.2312_:_SDS_page_and_coom SDS page and coomassie blue]

From an over night starter, cells were diluted and grown from Abs(600nm)=0.2 to Abs(600nm)=0.6.

Then 1UOD of cells (1.67ml at 0.6OD) was collected and centrifuged at 5000g for 5min.

After removal of the supernatant, the cell pellet was resuspended in 50µL SDS-PAGE sample buffer.

[http://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.2312_:_SDS_page_and_coom SDS page and coomassie blue] The mixture was loaded onto a polyacrylamide gel and migrated during 50min at 180V. Staining was done using coomassie blue.

The CsgA is at mass 19,6kDa, and can be clearly seen in the gel photograph.

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