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 20th September 1.1 Lab work 1.1.1 Visualization 1.1.1.1 Samples preparation for sequencing 1.1.1.2 NanoDrop Measurements 1.1.1.3 Gibson of cleaned up PCR products from pSB1C3 with dCas9 ST - GFP 11 (pPS16_017) clone 8 treated by DpnI Tuesday 20th September Lab work Visualization Samples preparation for sequencing "By Maxence, Mahnaz & Coline" 20 µL of plasmids pSB1C3 FKBP - GFP 10 (clones 7, 8, 9 and 10) were sent to be sequenced. 20 µL of the primers iPS84 (5µM) and iPS140 (5µM) were sent for sequencing. NanoDrop Measurements "By Maxence, Mahnaz & Coline" Sample Concentration (ng/µL) FKBP - GFP 10 clone 7 X FKBP - GFP 10 clone 8 X FKBP - GFP 10 clone 9 X FKBP - GFP 10 clone 10 X PCR product 1 obtained by iPS174 & iPS175 X PCR product 2 obtained by iPS173 & iPS176 X Gibson of cleaned up PCR products from pSB1C3 with dCas9 ST - GFP 11 (pPS16_017) clone 8 treated by DpnI By Maxence, Mahnaz & Coline Gibson was performed with cleaned up PCR products from pSB1C3 with dCas9 ST - GFP 11 (pPS16_017) clone 8 treated by DpnI, more precisely PCR product 1 obtained by iPS174 & iPS175 (plasmid) and PCR product 2 obtained by iPS173 & iPS176 (insert) with the following protocol: For each 20μl of reaction, mix the following reagents : 0.2 µL of insert 3.16 µL of plasmid 6.64 µL of water 10 µL of buffer mix Two controls were done: one without insert and one without buffer (water was added in order to have a total of 20 µL). The PCR was performed as follow : 1 hour at 50°C.
"By Maxence, Mahnaz & Coline"
20 µL of plasmids pSB1C3 FKBP - GFP 10 (clones 7, 8, 9 and 10) were sent to be sequenced. 20 µL of the primers iPS84 (5µM) and iPS140 (5µM) were sent for sequencing.
By Maxence, Mahnaz & Coline
Gibson was performed with cleaned up PCR products from pSB1C3 with dCas9 ST - GFP 11 (pPS16_017) clone 8 treated by DpnI, more precisely PCR product 1 obtained by iPS174 & iPS175 (plasmid) and PCR product 2 obtained by iPS173 & iPS176 (insert) with the following protocol:
For each 20μl of reaction, mix the following reagents :
Two controls were done: one without insert and one without buffer (water was added in order to have a total of 20 µL). The PCR was performed as follow : 1 hour at 50°C.
