Team:Groningen/Notebook/tasA Colony PCR t66
Blue Bio Energy
tasA Colony PCR (t66)
The colony PCR is performed to check the insert of the 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.
On gel it was seen that two samples of the colony PCR possibly had the insert.
Colony PCR
00:00, 9 August 2015 - 00:00, 9 August 2015
The transformation of 7 August was checked. Some white colonies were visible. With these colonies a colony PCR was carried out. After the PCR the samples were loaded on a 1% agarose gel and it was seen that sample 3 and 7 possibly have the insert. Cultures were made of sample 3 and 7 by putting some of the bacteria in a tube with 3 mL LB+cm. The tubes were put in the shaking incubator at 37 °C with 200 rpm.