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 Sunday 2nd October 1.1 Lab work 1.1.1 Visualization 1.1.1.1 Cloning of FRB - GFP 11 (pPS16_019) in FKBP - GFP 10 - pSB1C3 (pPS16_018) by digestion-ligation 1.1.1.2 Cloning of GFP 1.9 from pUC19 (pPS16_009) in pSB1C3 by digestion-ligation 1.1.1.3 Plasmids extraction of FRB - GFP 11 in pSB1C3 (pPS16_019) clone 4, FKBP - GFP 10 in pSB1C3 (pPS16_018) clone 6 and GFP 1.9 in pUC19 (pPS16_009) clone 1 Sunday 2nd October Lab work Visualization Cloning of FRB - GFP 11 (pPS16_019) in FKBP - GFP 10 - pSB1C3 (pPS16_018) by digestion-ligation By Maxence & Victor PRECIPITATION ETOH Once template and vector were cut, DNA ligase was used to join the sticky ends of the template and vector together: 1.5 µL of Buffer T4 10X 1 µL of ligase T4 enzyme 1 µL of ligase T4 enzyme 12.5 µL of water The mix were incubated for 30 minutes at rooming temperature. Cloning of GFP 1.9 from pUC19 (pPS16_009) in pSB1C3 by digestion-ligation By Maxence & Victor Once template and vector were cut, DNA ligase was used to join the sticky ends of the template and vector together: 6 µL of template (pPS16_009 treated by PstI & XbaI) 3 µL of vector (Bba treated by PstI & XbaI) 1.5 µL of Buffer T4 10X 1 µL of ligase T4 enzyme 3.5 µL of water The mix were incubated for 30 minutes at rooming temperature. Plasmids extraction of FRB - GFP 11 in pSB1C3 (pPS16_019) clone 4, FKBP - GFP 10 in pSB1C3 (pPS16_018) clone 6 and GFP 1.9 in pUC19 (pPS16_009) clone 1 "By Maxence & Victor" The following plasmids were extracted using the "Charge Switch-Pro Plasmid Miniprep" kit: pPS16_018 (FKBP - GFP 10) clone 6 pPS16_019 (FRB - GFP 11) clone 4 pPS16_009 (GFP 1.9) clone 1
By Maxence & Victor
PRECIPITATION ETOH
Once template and vector were cut, DNA ligase was used to join the sticky ends of the template and vector together:
The mix were incubated for 30 minutes at rooming temperature.
"By Maxence & Victor"
The following plasmids were extracted using the "Charge Switch-Pro Plasmid Miniprep" kit:
