Team:UCLA/Notebook/Recombinant Expression/5 August 2015
The two start up cultures are taken from the shaking incubator and the OD checks are 0.3 and 0.2, respectively. The start-up culture with the higher OD is poured into a L of LB with 1mL of Chloramphenicol. The contents are put into a shaking incubator at 37 degrees Celsius and left to incubate for 3hours. A 1mL sample is taken from the flask, and IPTG is added to a final concentration of 0.5mM. After IPTG addition, a 1mL sample of culture is taken every hour after IPTG addition for 4 hours total. The samples will be used in an SDS PAGE to check for verification of over expression of the 12-mer proteins. The contents are then left to incubate overnight for approximately 17 hours.
Observations Before IPTG addition, a 1mL sample of culture is extracted form the flask and pipetted into a 1.5mL micro centrifuge tube. The contents are spun with a balance at 3000 rpm for a minute. However, no visible cell pellet appeared at the bottom of the tube. It seemed like