User:Paulina Alatriste/Notebook/UNAM Genomics Mexico 2011/2011/07/28

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Plasmid Characterization-Stability, continuation

  • Continuation of the experiments in E. coli for measure plasmid stability: count of colonies, picked of new colonies, culture of new dilutions at hour 36.

  • Today the first thing that Pablo and I did was to check if there were colonies in the petri dishes we plated yesterday. There were colonies in the 6 petri dishes. So the next step was to pick colonies with a sterile stick and striate one little line in two petri dishes one with LB medium+gentamicin an the other with only LB media (all with the same stick). I picked 100 colonies of hour 0 and 100 colonies of hour 12 and I left the petri dishes incubating at 37°C.
  • At 9:20 pm I took another 1 mL of the stock culture (left it growing for 36 hours now) and repeat the following procedure:
1. Vortex the 1 mL of the stock culture.
2. Take 100μl of the previous culture and put them in 900 μL of liquid LB medium without antibiotic. This is the first dilution.
3. Vortex 30 times the dilution.
4. Repeat the steps 2 and 3 until reach the desired dilution.

I did dilutions 1010-5, 10-6, 10-7. An plated 100μL of each one in 3 petri dishes.