Tuesday 21^st June

Heat-shock competent cells preparation

By Lea, Caroline and Marion

Culture OD was measured at 600nm (OD_600nm = 3.4). 3.4mL were put into 250mL of LB medium and incubated at 20°C and 180rpm overnight.

Interlab study

Transformation (cf. protocole)

By Lea, Caroline and Marion

50µL of competent DH5α cells were transformed with 1µL of plasmids (for each construction: device 1, 2, 3, negative control and positive control) and one control tube was made without plasmids. Tubes were kept into ice for 30min then a thermal shock was made at 42°C for 1min. 500µL LB medium was added into each tube and incubated at 37°C for one 1h. Petri dishes were prepared with selective medium containing LB, agar and 30µg/mL chloramphenicol. Each transformation condition was spread on Petri dishes in duplicate and incubated overnight at 37°C.