User:Howard Boland/Notebook/Art from Synthetic Biology/2011/03/14

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BBa_katE colony screening

Since my overnight growth did not have GFP and the miniprep with digest came up with three bands (~3kb, ~9kb, ~2.1kb) it is likely that there has been some sort of self-ligation or DNA contamination. However, since the petridish show GFP colonies and I can verify these under the microscope I picked colonies and screened them under the microscope

For two labelled eppendorf tubes

  1. Add 10µl of H2O in each tube
  2. Transfer 1 separate colony to each of the two tubes
  3. Mix well
  4. Move 5µl of mix onto each of two slides, label
  5. Observe under microscope

I found one colony showing good GFP generation the other showed no GFP. I selected the GFP and pulled out the remaining 5µl from the tube and grew this overnight.