Protocols

Wet-lab on eGFP-expressiong E.Coli

During the semester, we went to the lab to manipulate the bacteria.

We made a Kanamycin-LB solution to only select the bacteria Dr. van der Meer gave us.

To get a 50mg/ml solution, we diluted 0.5g of solid kanamycin into 0.5ml of ddH2O (double distilled water).

The following day, we made serial dilutions of our overnight grown colonies to decrease the number of bacteria present in the solution, and make it easier to analyze them later.

http://faculty.irsc.edu/FACULTY/TFischer/micro/serial%20dilution.jpg

We then added 10ul of Kanamycin in the agar-LB plates we had prepared in a previous session. to let only our strains grow. The petri dishes contained 10ml of agar-LB. We then let the colonies grow to count them later.

We also made a glycerol stock to be able to store the bacteria at -80°C during a longer time. For that, we used:

• 0.5 ml of 40% gylcerol (it will prevent the water to expand while freezed, and preserve the bacteria)
• 0.5 ml of our sample bacteria (As & eGFP-only E.Coli)

We poured the glycerol in the vial first to avoid contamination, then gently vortex the solution and stored it on dry ice.

eGFP-expressing E.Coli strain (2676) expression map

Wet-lab at Dr. van der Meer's

Detailed Protocol here.

Arsenic-eGFP strain (1598) expression map