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 Friday 29th July 1.1 Lab work 1.1.1 Visualization 1.1.1.1 gBlock 1.1, 1.2 and GFP1-9 insertion in puc19 1.1.1.2 High fidelity PCR on bacteria transformed with pPS16_001, pPS16_002 and pPS16_005 Friday 29th July Lab work Visualization gBlock 1.1, 1.2 and GFP1-9 insertion in puc19 By Caroline The insertion was carried out following the usual protocol. High fidelity PCR on bacteria transformed with pPS16_001, pPS16_002 and pPS16_005 By Alice and Mathilde A PCR with Q5® High-Fidelity 2X Master Mix was performed on clones selected after screening PCR made on July 28, following this protocol.1151_pheoR and 1152_pheoF primers were used. Annealing temperature was 66°C and initial denaturation was 3 min. After amplification, 1 µL of loading dye was added to 5µL of each PCR products. Then 5 µL of this mix were put into a well of the gel. A last well contained 10µL of DNA ladder. PCR products were migrated 30min at 100V. PCR products expected were : Plasmids Band size (bp) pPS16_001 1017 pPS16_002 1017 pPS16_005 1017
By Caroline
The insertion was carried out following the usual protocol.
By Alice and Mathilde
A PCR with Q5® High-Fidelity 2X Master Mix was performed on clones selected after screening PCR made on July 28, following this protocol.1151_pheoR and 1152_pheoF primers were used. Annealing temperature was 66°C and initial denaturation was 3 min. After amplification, 1 µL of loading dye was added to 5µL of each PCR products. Then 5 µL of this mix were put into a well of the gel. A last well contained 10µL of DNA ladder. PCR products were migrated 30min at 100V.
PCR products expected were :
