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 29th August 1.1 Lab work 1.1.1 Visualization 1.1.1.1 PCR of the plasmid extraction from clonies 4,5,9,12 containing FRB sequence in pJET plasmid 1.1.1.2 NanoDrop Measurements Monday 29th August Lab work Visualization PCR of the plasmid extraction from clonies 4,5,9,12 containing FRB sequence in pJET plasmid By Mahnaz 2.5 µL DreamTaq Buffer 0.5 µL of dNTPs (10mM) 1 µL of each primer (10µM) primer pJET R and F 0.13 μl of DreamTaq Pol 20 µl H2O PCR was performed as follow: Step Temperature Time Initial denaturation 95°C 3 min 30 cycles 95°C 30 sec 52 30 sec 72°C 1 min Final Extension 72°C 7min Hold 4°C \infinity\ Primers used were: Matrix Clones containing dCas9 NM - GFP 10 in pSB1C3 Primers pJET R and F Tm 60°C t 1 min 30 After amplification, 3 µL of each PCR products and 5 µL of DNA ladder were placed in wells and migrated at 100V during 30 min. PCR products expected were : PCR products Expected band size (bp) FRB in pJET 474 GEL GEL GEL all 4 plasmids were sent for sequencing. NanoDrop Measurements By Mahnaz Sample Concentration (ng/µL) PCR fragment FRB clone 4 58.43 PCR fragment FRB clone 5 320.75 PCR fragment FRB clone 9 830.99 PCR fragment FRB clone 12 473.37
By Mahnaz
PCR was performed as follow:
Primers used were:
After amplification, 3 µL of each PCR products and 5 µL of DNA ladder were placed in wells and migrated at 100V during 30 min.
PCR products expected were :
GEL GEL GEL
all 4 plasmids were sent for sequencing.
