Toggle navigation Home Team Team Media Collaborations Sponsors Acknowledgements Project Background Design CRISPR/Cas9 Strategy Experiments Notebook Results Perspective Interlab Study Parts Parts Basic Parts Composite Parts Human Pratices Overview Societal Issues of CRISPR/Cas9 Responsible Research and Innovation GMO regulation Integrated Practices Engagement Model Attributions Safety Contents 1 Tuesday 21st June 1.1 Heat-shock competent cells preparation 1.2 Interlab study 1.2.1 Transformation (cf. protocole) Tuesday 21st June Heat-shock competent cells preparation By Lea, Caroline and Marion Culture OD was measured at 600nm (OD600nm = 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.
By Lea, Caroline and Marion
Culture OD was measured at 600nm (OD600nm = 3.4). 3.4mL were put into 250mL of LB medium and incubated at 20°C and 180rpm overnight.
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.
