From 2007.igem.org

Miniprep of pKD46

It will be used for the wanner procedure to recombine thermo-dependently

Purification of the wanner PCR of L58

We got a bright band to about 1,7kb on gel, and some higher non-specific products.
the 1,7kb band has been cuted out from the gel and purificated.

Transformation of W121 and FR781 with pKD46

We transformed the strains W121 and FR781, which are both DAP-, with the wanner recombinase plasmid pKD46.
1µL of the miniprep of the day has been used on 100µL of competent TSS-made cells.
Expression step and plate step are done at 30°C considering the thermosensibility of the plasmid.