"

From 2014.igem.org

Sentivity to antibiotics

After having succeeded to make our bacteria grow up, we tested their resistance to different antibiotics. To know their resistance to antibiotics is very important, because it will allow us to finalize protocols of selection after tranformation.
We chose to test the most commonly used antibiotics. We included the three antibiotics used in iGEM (Kanamycin, Chloramphenicol and Ampicilin), plus the Erythromycin and the Tetracyclin. We chose the erythromycin to test a conjugation protocol which required this antibiotic for E.Coli, and we chose to test tetracyclin because it is quite often used for inducibles systems.


We tested several concentrations of each antibiotics (cf: Table 1), and we add a supplementary concentration for erythromycin because it is known to be not very effective on GRAM- bacteria.

The sensitivity tests were performed in four differents conditions (cf: Figure 1). We tested on Marine Broth(MB) 1X plates, MB 0.5X plates (used for conjugation) and M9 1X (+3% NaCl). We aslo tested in liquid M9 1X (+3% NaCl) to have a test in liquid culture, but unfortunately we cannot test on liquid MB cultures because this media is no usable for the spectrometer, so we cannot have a good information about the cells' growth.