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 Lab work 1.1.1 Visualization 1.1.1.1 Colony PCR 1.1.1.2 Cell Cultures Tuesday 23th August Lab work 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.
