Team:NTNU-Trondheim/Notebook/October

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Trondheim iGEM 2013

[https://2013.igem.org/Team:NTNU-Trondheim/TransformedBricks Transformed BioBricks]

Wednesday 02.10.2013

SDS-PAGE of bacterial samples with the tat_GFP_RFP (ER1) and tat_ProteinG construct

In order to determine if Protein G is produced in the S3-2B sample and wheater a GFP-RFP dimer is produced in the ER1 sample, a SDS-PAGE was performed. Liquid cell culture with ER1 (tat_GFP_RFP construct), S3-2B (tat_ProteinG construct) and wildtype ER2566 ''E.coli'' cells was centrifuged and the pallet was mixed with SDS loading buffer. This mix was then heated at 95 °C for 15 minutes. The SDS-PAGE results can be viewed in the figure below

Figure 1.

[[file:SDS-PAGE_bacterias.jpg|400px]] Figure: SDS-PAGE of bacterial samples. From the left: wild type ER2566, tat_GFP_RFP and tat_ProteinG There is evidently produced Protein G in the S3-2B sample as there is an additional band at the expected size of about .... kDa. There is also an additional band at the ER1 sample of about ..... kDa. This half the expected size of about 60 kDa. This indicates that only RFP is produced in the bacterias.