Template:IONIS Paris/Notebook/15-07-15
Test of competent cells (transformation with pUC19)
Transformation of 50µl of competent cells with 1 µl of pUC19 following our protocol
50µL of competent cells have been used as negative control
Both of them have been plated onto agar plates containing chloramphenicol
Digestion of Gibson product (VVD-YC155)
NEB double digestion: Buffer 2.1
EcoRI:100%, PstI:75%
| Tube | Gibson |
|---|---|
| Water | 12 µL |
| Buffer 2.1 | 2 µL |
| Plasmid | 5 µL |
| Enzyme EcoRI | 0,5 µL |
| Enzyme PstI | 0,5 µL |
37°C, 60 min
Electrophoresis of digested products
Expected results
Results
We get expected bands at approximatively 2000 bp and 750 bp, but it still an extra band at 1500 bp (already present during the previous electrophoresis). That could correspond to another plasmid.
15 July 15