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 22nd July 1.1 Visualization 1.1.1 Low Fidelity Dreamtaq PCR of DH5α|pPS16_002 1.1.2 High Fidelity Q5 PCR of transformed DH5α with pPS16_004 and pPS16_007 1.1.3 Glycerol stocks for DH5α transformed with pcl TAA, TAG and Tq Friday 22nd July Visualization Low Fidelity Dreamtaq PCR of DH5α|pPS16_002 By Mathilde A DreamTaq PCR was made with DH5α|pPS16_002 cultures following the usual protocol with Tm at 57°c and 5min for the initial denaturation. We divided up the PCR mix in 6 PCR tubes and added in each one a different clone from the transformed culture. The picked colonies were re-plated on a Petri dish with LB + Ampicillin (50µg/mL) + X-Gal/IPTG (1/1000). Results : PCR products expected were : Plasmid pPS16_002 Band Size (bp) 960 The electropheresis on agarose gel showed absolutely no PCR products. pPS16_002 transformation from the 19/07/2016 is re-plated (50µL) on two Petri dishes LB + Ampicillin (50µg/mL) + X-Gal/IPTG (1/1000). High Fidelity Q5 PCR of transformed DH5α with pPS16_004 and pPS16_007 By Laetitia The PCR was performed following the usual protocol. 8 tubes were done: 5 clones of pPS16_004 and 3 clones of pPS16_007 The TM was at 60°C Glycerol stocks for DH5α transformed with pcl TAA, TAG and Tq By Laetitia 8 stocks were made: 2 clones (Cl 1 and Cl 2) : Cl1 and Cl2 of pcl TAA Cl1 and Cl2 of pcl TAG Cl1 and Cl2 of pcl Tq For 1 glycerol stock: 1mL of liquid culture 500 μL of glycerol 60%
By Mathilde
A DreamTaq PCR was made with DH5α|pPS16_002 cultures following the usual protocol with Tm at 57°c and 5min for the initial denaturation.
We divided up the PCR mix in 6 PCR tubes and added in each one a different clone from the transformed culture. The picked colonies were re-plated on a Petri dish with LB + Ampicillin (50µg/mL) + X-Gal/IPTG (1/1000).
Results : PCR products expected were :
The electropheresis on agarose gel showed absolutely no PCR products. pPS16_002 transformation from the 19/07/2016 is re-plated (50µL) on two Petri dishes LB + Ampicillin (50µg/mL) + X-Gal/IPTG (1/1000).
By Laetitia
The PCR was performed following the usual protocol. 8 tubes were done: 5 clones of pPS16_004 and 3 clones of pPS16_007
The TM was at 60°C
8 stocks were made: 2 clones (Cl 1 and Cl 2) :
For 1 glycerol stock:
