Team:Groningen/Notebook/tasA Transformation t47
Blue Bio Energy
tasA Heat shock transformation (t47)
The ligation product of tasA, Psalt and K823023 was transformed into E. coli.
Our goal is to overexpress TasA by adding a second copy of the tasA gene under the control of a salt inducible promoter. This overexpression will lead to a stronger biofilm when the biofilm is exposed to salt water.
The heat shock transformation was performed.
Heat shock transformation
00:00, 16 July 2015 - 00:00, 16 July 2015
Afterwards the heat shock transformation from the protocol book was performed. For this, 10 µL competent NEB cells with 10 µL ligation product was used. The resulting cells were plated on LB+amp plates and grown overnight at 37 °C.
00:00, 17 July 2015 - 00:00, 17 July 2015
The transformation of the ligation product of the salt promoter with tasA was checked. On the plates a few white colonies were seen. Miniprep and colony PCR were performed to check whether the insert size was the right one. The colony PCR showed possible tasA with Psalt insert in one sample. This colony was grown overnight in LB+amp.