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 5th July 1.1 Lab work 1.1.1 Visualization 1.1.1.1 Transformation of DH5α|pPS16_004 and DH5α|pPS16_007 1.1.2 Biobrick characterization 1.1.2.1 B-Galactosidase and luciferase test on transformed BL21 Tuesday 5th July Lab work Visualization Transformation of DH5α|pPS16_004 and DH5α|pPS16_007 By Caroline and Mathilde The 04/06/2016 transformations show white colony growth for gBlocks 4.2 clone 3 and GFP1-9 (clone 1), but blue colonies only for 4.1 clone 3. Thus this last gBlock 4.1 clone 3 was transformed again following the same protocol as the 21/06/2016 with : 50μg of competent DH5α cells 4.1 ligation product in 5μL of the plasmid puc19 4.1 clone 6 extracted DNA (which presented a good size strip on electrophoresis) 6 clones of each of the other transformations pPS16_001, pPS16_002, pPS16_003, pPS16_005 and pPS16_006 were grown in 4mL of LB with Ampicillin (50µg/mL) at 37°C, 180rpm. For pPS16_002 only 2 clones were cultivated because there was not more colony. To obtain more colonies, another transformation was made with 50µL of cells and 5µL of plasmid. 50µL of transformed bacteria were plated on LB + Ampicillin (50µg/mL). Biobrick characterization B-Galactosidase and luciferase test on transformed BL21 By Charlene Cultures tested Plasmid(s) K1372001 K1372001 + pcl UAA K1372001 + pcl UAG K1372001 + pcl Tq Clone 1 2 1 2 1 2 1 2 1 2 1 2 1 2 1 2 1 2 1 2 1 2 1 2 Salicylate concentration 0 30µM 1mM 0 30µM 1mM 0 30µM 1mM 0 30µM 1mM Ratio between luciferase activity and β-Gal activity
By Caroline and Mathilde
The 04/06/2016 transformations show white colony growth for gBlocks 4.2 clone 3 and GFP1-9 (clone 1), but blue colonies only for 4.1 clone 3.
Thus this last gBlock 4.1 clone 3 was transformed again following the same protocol as the 21/06/2016 with : 50μg of competent DH5α cells 4.1 ligation product in 5μL of the plasmid puc19 4.1 clone 6 extracted DNA (which presented a good size strip on electrophoresis)
6 clones of each of the other transformations pPS16_001, pPS16_002, pPS16_003, pPS16_005 and pPS16_006 were grown in 4mL of LB with Ampicillin (50µg/mL) at 37°C, 180rpm. For pPS16_002 only 2 clones were cultivated because there was not more colony. To obtain more colonies, another transformation was made with 50µL of cells and 5µL of plasmid. 50µL of transformed bacteria were plated on LB + Ampicillin (50µg/mL).
By Charlene
Cultures tested
