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 Tuesday 5th July Lab work Visualization Transformation of DH5α|pPS16_004 and DH5α|pPS16_007 The 04/06/2016 transformations show white colony growth for G-blocks 4.2 clone 3 and GFP1-9 (clone 1), but blue colonies only for 4.1 clone 3. Thus this last G-block 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 placed in 4mL of LB added to Ampicillin (50µg/mL) at 37°C, 180RPM. For 2.2 just 2 clones were cultivated because there was not more colony.
The 04/06/2016 transformations show white colony growth for G-blocks 4.2 clone 3 and GFP1-9 (clone 1), but blue colonies only for 4.1 clone 3.
Thus this last G-block 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 placed in 4mL of LB added to Ampicillin (50µg/mL) at 37°C, 180RPM. For 2.2 just 2 clones were cultivated because there was not more colony.
