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 Wednesday 27th July 1.1 Lab work 1.1.1 Visualization 1.1.1.1 Transformation of DH5a cells 1.1.2 Biobrick Characterization 1.1.2.1 BL21 electrocompetent cells preparation for glycerol preparation Wednesday 27th July Lab work Visualization Transformation of DH5a cells By Léa DH5a were transformed with pPS16_001, pPS16_002, pPS16_005, or pPS16_009 using the usual protocol. pPS16_002 ligation products of the 19th of July and the 27th of July were used. pPS16_001, pPS16_005 and pPS16_009 ligation products of the 26th of July were used. The Heat shock were performed at 42°C during 60 seconds. After a one hour incubation, transformation products were streaked on Petri dishes containing Ampicilin, X-Gal and IPTG, and incubated at 37°C overnight. Biobrick Characterization BL21 electrocompetent cells preparation for glycerol preparation By Charlène We obtained transformed bacteria for every conditions made yesterday (except for controls). Colonies of BL21 transformed with pcl_TAA and K1372001 Colonies of BL21 transformed with pcl_TAG and K1372001 Colonies of BL21 transformed with pcl_Tq and K1372001 We wanted to have glycerol stock of our transformed bacteria so we had to make a pre-culture. 3 clones of each condition were put in 2mL of LB + Streptomycin (50µg/mL) + Chloramphenicol (30µg/mL) medium. They were incubated overnight at 37°C, 180 rpm.
By Léa
DH5a were transformed with pPS16_001, pPS16_002, pPS16_005, or pPS16_009 using the usual protocol. pPS16_002 ligation products of the 19th of July and the 27th of July were used. pPS16_001, pPS16_005 and pPS16_009 ligation products of the 26th of July were used. The Heat shock were performed at 42°C during 60 seconds.
After a one hour incubation, transformation products were streaked on Petri dishes containing Ampicilin, X-Gal and IPTG, and incubated at 37°C overnight.
By Charlène
We obtained transformed bacteria for every conditions made yesterday (except for controls).
We wanted to have glycerol stock of our transformed bacteria so we had to make a pre-culture. 3 clones of each condition were put in 2mL of LB + Streptomycin (50µg/mL) + Chloramphenicol (30µg/mL) medium. They were incubated overnight at 37°C, 180 rpm.
