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 30th august 1.1 Visualization 1.1.1 Plasmids extraction 1.1.2 Gibson Assembly of segments 1 and 2 1.1.3 Transformation in DH5α with the 1-2 Gibson assembly product Tuesday 30th august Visualization Plasmids extraction "By Mahnaz" Plasmid pZA11 were extracted using the "Charge Switch-Pro Plasmid Miniprep" kit Gibson Assembly of segments 1 and 2 By Mahnaz Two preparations were made : the 1-2 Gibson assembly product tube : 1,12µL of purified segment 1 and 0,56µL of purified segment 2 + 0,85µL of digested pSB1C3 + 7,47 µL of sterile water + 10µL of NEB Builder Hifi DNA Assembler Master Mix the control tube : 0,85µL of digested pSB1C3 + 17,47 µL of sterile water Gibson products were incubated 1h at 52°c. Transformation in DH5α with the 1-2 Gibson assembly product By Mahnaz The transformation was performed following the usual protocol with 2µL of plasmid. 50, 150 and 350µL of the transformed DH5α with the 1-2 Gibson assembly product were plated on 3 plates of LB + chloramphenicol (30µg/mL) 350 µL of each Gibson assembly control and transformation control were plated on 2 plates of B + chloramphenicol (30µg/mL) Petri cultures were incubated at 37°c overnight.
"By Mahnaz"
Plasmid pZA11 were extracted using the "Charge Switch-Pro Plasmid Miniprep" kit
By Mahnaz
Two preparations were made :
Gibson products were incubated 1h at 52°c.
The transformation was performed following the usual protocol with 2µL of plasmid.
Petri cultures were incubated at 37°c overnight.
