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 Monday 27th June 2 Lab Meeting 2.1 Labwork 2.1.1 Interlab study 2.1.1.1 Cell culture 2.1.2 Visualization 2.1.2.1 BioBrick construction 2.1.3 Bringing DNA closer 2.1.3.1 Cell culture 2.1.4 BioBrick characterization Monday 27th June Lab Meeting Members present: Instructors and advisors: Claire, Fabio, Marie, Olivier, Sylvie, Philippe. Students: Carla, Caroline, Charlène, Claire, Coline, Léa, Mahnaz, Marion, Martin, Maxence, Naiane, Yacine Labwork Interlab study Cell culture By Lea Transformed bacteria frozen on 24/06/2016 were put into 3mL of LB medium containing 30µg/mL chloramphenicol and incubated overnight at 37°C, 180rpm. Visualization BioBrick construction By Caroline, Charlène and Naïane GBlocks from IDT were received on the 24/06/2016. pUC19 plasmid was digested using HincII restriction enzyme. Component Volume (µL) Plasmid 5 Tango buffer 1x 5 Water 38 Enzyme 1 The mix was incubated for 1 hour at 37°C. An electrophoresis was done (0.8% of agarose). Component Volume (µL) Digested DNA 5 Water 5 Loading buffer 2 A DNA-ladder was used to visualize the DNA fragments size. Bringing DNA closer Cell culture By Alice and Lea 4 plasmids containing NM Cas9 (DS-NMcas), SP Cas9 (DS-SPcasN-), ST1 Cas9 (DS-ST1casN-) and Td Cas9 (DS-TDcasN-) were ordered to Addgene and received on 24/06/2016. Plasmids were delivered into tubes containing LB agar medium and bacteria colony containing plasmids. Bacteria were put into 15mL of LB medium containing 50µg/mL of spectinomycin and incubated overnight at 37°C, 180 rpm. BioBrick characterization By Alice and Lea BioBrick [http://parts.igem.org/Part:BBa_K1372001 K1372001] from iGEM team Paris Saclay 2014 was chosen to be characterized. 10µL of DH5α|K1372001 cells were put into 3mL of LB medium containing 30µg/mL of chloramphenicol and incubated overnight at 37°C, 180 rpm.
Members present:
By Lea
Transformed bacteria frozen on 24/06/2016 were put into 3mL of LB medium containing 30µg/mL chloramphenicol and incubated overnight at 37°C, 180rpm.
By Caroline, Charlène and Naïane
GBlocks from IDT were received on the 24/06/2016.
pUC19 plasmid was digested using HincII restriction enzyme.
The mix was incubated for 1 hour at 37°C. An electrophoresis was done (0.8% of agarose).
A DNA-ladder was used to visualize the DNA fragments size.
By Alice and Lea
4 plasmids containing NM Cas9 (DS-NMcas), SP Cas9 (DS-SPcasN-), ST1 Cas9 (DS-ST1casN-) and Td Cas9 (DS-TDcasN-) were ordered to Addgene and received on 24/06/2016. Plasmids were delivered into tubes containing LB agar medium and bacteria colony containing plasmids. Bacteria were put into 15mL of LB medium containing 50µg/mL of spectinomycin and incubated overnight at 37°C, 180 rpm.
BioBrick [http://parts.igem.org/Part:BBa_K1372001 K1372001] from iGEM team Paris Saclay 2014 was chosen to be characterized. 10µL of DH5α|K1372001 cells were put into 3mL of LB medium containing 30µg/mL of chloramphenicol and incubated overnight at 37°C, 180 rpm.
