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 23th August 1.1 Visualization 1.1.1 Colony PCR 1.1.2 Cell Cultures Tuesday 23th August Visualization Colony PCR By Léa and Naiane 12 colonies containing the gBlock GFP 1-9 and 12 colonies transformed with Gibson Assembly products (fragment 3-4) were screened and used for the usual protocol of Colony PCR. The same colonies were also plated on Petri dish containing solid LB + Cm and liquid culture (LB + Cm) was made from these same colonies. Both petri dishes and the liquid cultures were overnight at 37°C. The migration on agarose gel was done the next day. Cell Cultures By Léa, Naiane and Mathilde 100µL ofDH5a precultures containing dCas9 Nm plasmids (clone 1 and 2) were used to inoculate 200mL of liquid LB medium containing spectinomycin. Interlab study plates from the 22/08/2016 were used to inoculate two tubes containg 5mL of liquid LB medium (chloramphenicol) per construction. DH5a colonies containing pUC19 vector cloned with sgRNA NM (clone 5 and 12) or 1.2 (clones 8 and 14) were inoculated into 4mL of liquid LB medium.
By Léa and Naiane
12 colonies containing the gBlock GFP 1-9 and 12 colonies transformed with Gibson Assembly products (fragment 3-4) were screened and used for the usual protocol of Colony PCR. The same colonies were also plated on Petri dish containing solid LB + Cm and liquid culture (LB + Cm) was made from these same colonies. Both petri dishes and the liquid cultures were overnight at 37°C. The migration on agarose gel was done the next day.
By Léa, Naiane and Mathilde
100µL ofDH5a precultures containing dCas9 Nm plasmids (clone 1 and 2) were used to inoculate 200mL of liquid LB medium containing spectinomycin.
Interlab study plates from the 22/08/2016 were used to inoculate two tubes containg 5mL of liquid LB medium (chloramphenicol) per construction.
DH5a colonies containing pUC19 vector cloned with sgRNA NM (clone 5 and 12) or 1.2 (clones 8 and 14) were inoculated into 4mL of liquid LB medium.
