Ligation FC

Screening of the cloning of E0240 and FlhDC+promotor

Spreading the clones in order to obtain single colonies

The PCR screening of the transformants L139 and L142 of august 15th revealed several bands for a given clone including one band appearing at the right size.
There are 2 hypothesis:

• The right clone was contaminated by a wrong one
• The clone contains 2 plasmids: one with the insert and another one without the insert

In order to check these 2 hypothesis and to isolate (if it is possible) the right clone (containing the plasmid with the insert). We decided to spread the "clone" in question in a LB plate in order to carry out a PCR screening on single colonies.

• Take of some bacteria from the glycerol stock
• Resuspension in 400 µL of LB+antibiotic
• Spreading of 200 µL in a LB agar plate containing the appropriate antibiotic
• Incubation overnight at 37°C