- Circularization of previously digested plasmids from which the tag of periplasm exportation was extracted.
- Helena previously cut HydA and PFOR1 plasmids with NdeI in order to eliminate the tag of periplasm exportation from the plasmids where HydA and PFOR1 parts are inserted. We need to circularize again this fragments to perform a PCR and amplify HydA and PFOR1 regions. So today I made 5 ligation reactions with the following proportions:
|Buffer 10x T4 Ligase
- I left the tubes incubating at 16°C for 12 hours
I made two reactions to circularize HydA and 3 to circularize PFOR1.