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 11th July 1.1 Lab work 1.1.1 Biobrick characterization 1.1.1.1 Electrocompetents and transformation by electroporation of BL21 1.1.2 Bringing DNA closer 1.1.2.1 DH5α transformation with DS-NMcas, DS-SPcasN-, DS-ST1casN- and DS-TDcasN- Monday 11th July Lab work Biobrick characterization Electrocompetents and transformation by electroporation of BL21 By Charlène BL21 cells were grown up to OD600nm=7.14. They were then made competent and transformed with K1372001 extracted from clone 2 and pcl_TAA, pcl_TAG or pcl_Tq to create the following strains: BL21|K1372001+pcl_TAA BL21|K1372001+pcl_TAG BL21|K1372001+pcl_Tq Bringing DNA closer DH5α transformation with DS-NMcas, DS-SPcasN-, DS-ST1casN- and DS-TDcasN- By Laetitia and Mathilde DS-NMcas, DS-SPcasN-, DS-ST1casN- and DS-TDcasN- plasmids were resuspended with 5μL of sterilized water. 5 tubes were prepared : one control tube with only 50μL of DH5α heat-shock competent cells four tubes with 50μL of DH5α heat-shock competent cells + 1μL of plasmid The regular heat-shock competent cells transformation protocol was used. Transformed cells were plated in duplicata (10μL and 50μL) on 8 Petri dishes (spectinomycin 50μg/mL). Control cells were plated on another plate (spectinomycin 50μg/mL).
By Charlène
BL21 cells were grown up to OD600nm=7.14. They were then made competent and transformed with K1372001 extracted from clone 2 and pcl_TAA, pcl_TAG or pcl_Tq to create the following strains:
By Laetitia and Mathilde
DS-NMcas, DS-SPcasN-, DS-ST1casN- and DS-TDcasN- plasmids were resuspended with 5μL of sterilized water. 5 tubes were prepared :
The regular heat-shock competent cells transformation protocol was used.
Transformed cells were plated in duplicata (10μL and 50μL) on 8 Petri dishes (spectinomycin 50μg/mL). Control cells were plated on another plate (spectinomycin 50μg/mL).
