ORBIT: Protocols

ORBIT is a complete reverse genetic toolkit, therefore the types of modifications that can be made vary and should be taken into consideration when planning the initial ORBIT experiment. This flowchart provides an overview of how to approach an ORBIT experiment depending on whether the strain you are making will retain the entire integrating plasmid or have part (or all) of it removed (markerless / scarless modification).

The detailed protocols below walk through how to make induced electrocompetent cells and perform ORBIT integrations. Combining these protocols with the overview above should enable many avenues of strain construction.