Team:Northwestern/OMV purification

Adapted Extracellular Vesicle preparation protocol:

• Grow desired volume of E. coli liquid culture until mid-log phase (OD₆₀₀ = 0.5-0.8)
• Take note of exact OD₆₀₀
• Pellet the cells gently (approx. 6,000 g) for 5 minutes
• Recover as much supernatant as possible without disturbing the cell pellet
• Leave some supernatant behind if necessary
• Take note of the volume recovered
• Filter the supernatant using a 0.22 µm syringe filter unit to remove any remaining cells
• Change unit if and whenever it clogs
• Equilibrate a 14 kDa centrifugal filter unit with PBS
• Concentrate the filtered supernatant with the centrifugal filter unit following manufacturer’s instructions
• Once all the supernatant has been concentrated, if desired, buffer exchange using the same centrifugal filter unit (e.g. with PBS prior to a SDS-PAGE)
• Sample is ready for analysis.