- Tuesday: Transforming Competent cells
* We added our suspended DNA and competent cells together. * We added 200 micro liters of SOC broth. * We plated a total of 11 plates, 3 controls, and 2 different concentrations of DNA per BioBrick. * We then incubated overnight at 37°C.
- Wednesday: Collecting Colonies
* Josh counted all our colonies on the plates, all of our BioBricks grew except, BBa_K123002 * We got expected results with no colonies growing on the ampicilin plate. * Josh then added 5mL LB medium into labeled 50mL vials. * Josh selected and numbered 4 colonies from each part. * Josh then swabbed each of the numbered colonies and put the swab in its corresponding tube.
-Thursday: Plasmid DNA Preparation and storage
* We added 830μL of our LB growth medium mixture tubes to 320μL glycerol, we then stored in -80°C freezer. * Next we put 2mL from our LB growth medium mixture into 2mL tubes. * We then centrifuged the tubes. * We then dumped out the LB and repeated previous two steps. Thus leaving behind cells at the bottom. * We then used the Fermentas GeneJet plasmid miniprep kit to prepare our plasmid DNA. * We altered the last step by only adding 35μL of the Elution Buffer, then let it incubate, and then centrifuged for two min. We then repeated this process again. * We then stored the run through in -80°C freezer.