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 Monday 10th October 1.1 BioBrick K2039000 and K2039001 characterization 1.1.1 Protein extraction of FRB - GFP 11 in pSB1C3 (pPS16_019) clone 4, FKBP - GFP 10 in pSB1C3 (pPS16_018) clone 6, GFP 1.9 in pUC19 (pPS16_009) clone 1 and PhB1040 strain 594 1.1.2 Protein electrophoresis of FRB - GFP 11 in pSB1C3 (pPS16_019) clone 4, FKBP - GFP 10 in pSB1C3 (pPS16_018) clone 6, GFP 1.9 in pUC19 (pPS16_009) clone 1 and PhB1040 strain 594 1.1.3 Transfert of protein extracted from FRB - GFP 11 in pSB1C3 (pPS16_019) clone 4, FKBP - GFP 10 in pSB1C3 (pPS16_018) clone 6, GFP 1.9 in pUC19 (pPS16_009) clone 1 and PhB1040 strain 594 Monday 10th October BioBrick K2039000 and K2039001 characterization Protein extraction of FRB - GFP 11 in pSB1C3 (pPS16_019) clone 4, FKBP - GFP 10 in pSB1C3 (pPS16_018) clone 6, GFP 1.9 in pUC19 (pPS16_009) clone 1 and PhB1040 strain 594 By Maxence In order to perform a Western Blot, the proteins were extracted from clones (pPS16_018 clone 6, pPS16_019 clone 4, pPS16_009 clone 1 and PhB1040 strain 594) put on liquide culture the 7th October. For that purpose, the protein extraction protocol was used: Sample OD (1 ml) Volume of lysis buffer FRB - GFP 11 in pSB1C3 (pPS16_019) 5.1 127.5 μl FKBP - GFP 10 in pSB1C3 (pPS16_018) 5.2 130 μl GFP 1.9 in pUC19 (pPS16_009) 5.3 132.5 μl PhB1040 strain 594 2.8 70 μl Protein electrophoresis of FRB - GFP 11 in pSB1C3 (pPS16_019) clone 4, FKBP - GFP 10 in pSB1C3 (pPS16_018) clone 6, GFP 1.9 in pUC19 (pPS16_009) clone 1 and PhB1040 strain 594 By Maxence A protein electrophoresis of the extracted protein was done using the protein electrophoresis protocol. For that purpose, the gel cassette Bolt 4-12% Bis-Tris Plus 10W was used and the power was set for 1 hour at 65V. Transfert of protein extracted from FRB - GFP 11 in pSB1C3 (pPS16_019) clone 4, FKBP - GFP 10 in pSB1C3 (pPS16_018) clone 6, GFP 1.9 in pUC19 (pPS16_009) clone 1 and PhB1040 strain 594 By Maxence The proteins on the electrophoresis gel were transfered on blotting membrane using the protein transfert protocol.
By Maxence
In order to perform a Western Blot, the proteins were extracted from clones (pPS16_018 clone 6, pPS16_019 clone 4, pPS16_009 clone 1 and PhB1040 strain 594) put on liquide culture the 7th October. For that purpose, the protein extraction protocol was used:
A protein electrophoresis of the extracted protein was done using the protein electrophoresis protocol. For that purpose, the gel cassette Bolt 4-12% Bis-Tris Plus 10W was used and the power was set for 1 hour at 65V.
The proteins on the electrophoresis gel were transfered on blotting membrane using the protein transfert protocol.
