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 Thursday 21th July 1.1 Lab work 1.1.1 Biobrick characterization 1.1.1.1 K1372001 from clone 2 digestion 1.1.1.2 Making Petri dishes of medium LB (with strepto or IPTG and Xgal 1.1.1.3 Liquid culture of pcl TAA, TAG and Tq 1.1.2 Visualization 1.1.2.1 1.1.3 Get DNA Closer 1.1.3.1 Thursday 21th July Lab work Biobrick characterization K1372001 from clone 2 digestion By Alice and Terrence We made a digestion of K1372001 plasmid following this protocol. We used EcoR1 and Pst1 fast digest enzymes. After incubation, 3.3 mL of loading dye is added to digestion products. We put in wells 10µL of DNA ladder and 20 µL of digestion products. We set the eletrophoresis machine at 100V for 25min. Digestion products expected were : Band size (Bp) Plasmid 1590 Insert 2029 Migration of digested product of K1372001. Making Petri dishes of medium LB (with strepto or IPTG and Xgal By Laetitia A Petri dish with LB and streptomycin was made with: 20 mL of LB agar 10µL of streptomycin (initial concentration 100mg/mL) We added IPTG and Xgal on 4 petri dishes of LB and Ampicilin. For each petri dish: 500µL of water 1µL of IPTG 1µL of Xgal The solution was spreaded on petri dish Liquid culture of pcl TAA, TAG and Tq By Laetitia 2 clones of each petri dish were soaked in a tube of LB and Streptomycin. 6 tubes were made. For each tube: 1 mL LB 0.5 µL streptomycin The tubes were incubated at 37°C and 180 rpm overnight. Visualization By Get DNA Closer By
By Alice and Terrence
We made a digestion of K1372001 plasmid following this protocol. We used EcoR1 and Pst1 fast digest enzymes. After incubation, 3.3 mL of loading dye is added to digestion products. We put in wells 10µL of DNA ladder and 20 µL of digestion products. We set the eletrophoresis machine at 100V for 25min.
Digestion products expected were :
By Laetitia
A Petri dish with LB and streptomycin was made with:
We added IPTG and Xgal on 4 petri dishes of LB and Ampicilin. For each petri dish:
The solution was spreaded on petri dish
2 clones of each petri dish were soaked in a tube of LB and Streptomycin.
6 tubes were made. For each tube:
The tubes were incubated at 37°C and 180 rpm overnight.
By
