Team:Paris Saclay/Notebook/July/12
Contents
Monday 12th July
Lab work
Getting DNA closer
Amplification of DS-TDcasN, DS-SPcasN and pZA21 plasmide
By Caroline
Plasmids extracted the 4/07/16 were used to amplify the dCas9 needed for the get DNA closer tool and the pZA21 plasmid in which the tool will be inserted. A PCR was carry out following PCR with Q5® High-Fidelity 2X Master Mix from NEB protocol (cf protocols section) adapted to have 50µL at the end.
Biobrick characterization
Electrocompetents and transformation by electroporation of BL21
By Charlene
Cells transformed the 11/07/16 have grown all the night. However no colony have been observed. To control if there is a bacteria transformed which have been not putted on Petri dish, we performed another experiment. 100 µl (500µl concentrated) of transformed bacteria for each condition, conserved at -4°C from yesterday, have been split in 3 solutions : - 30µl of bacteria + 3ml LB + Streptomycin (50µg/ml) + Chloramphenicol (30µg/ml) - 30µl of bacteria + 3ml LB + Streptomycin (50µg/ml) - 30µl of bacteria + 3ml LB + Chloramphenicol (30µg/ml) They incubated 8h at 37°C, 200 RPM.
No bacteria have grown so we determinated that it was a problem of transformation.
