840:153g:Projects/project13/2010/09/23

From OpenWetWare
Jump to: navigation, search

- 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.