Get things ready

You will need these items for the experiment.

Items required for the experiment: Along with these, ensure to have sterile serological pipettes and a pipette controller available.

Hardware

• BactoBox® setup including a flow cell with at least 10 measurements.

• Windows 10/11 computer.

• 10 × shake flasks with vented caps. Preferably 50/125 mL capacity. Alternative: 2 × 6-deepwell microplates.

• Incubator shaker that can hold 10 flasks (not shown).

• Permanent marker.

• Vortex mixer.

• Pipettes and sterile pipette tips: P10,000, P1000, P200. Use extended pipette tips to reduce contamination risks.

• Sterile, serological pipettes. Preferably 10 mL version.

• Serological pipet controller (not shown).

• Sterile 15 mL centrifuge vial (or similar) for disaggregating overnight culture.

• Rack for 15 mL centrifuge tubes.

Reagents

• 1 × cryo stock of your bacterial species.

• Growth medium. Preferably filtered through 0.2 µm filter to minimize contribution of background particulates from growth medium.

• Dilution vials either ready-to-use or freshly prepared using the dilution dispenser.

• Disinfection vials.

Use proper aseptic technique

In the coming videos, we use a simple lab bench for the procedure. Always use proper aseptic techniques, including sterile pipette tips, laminar air flow (LAF), and disinfection of the work environment.

Fill level for different vessels.

• 50/125 mL shake flasks or 6-deepwell microplates: Add 9 mL cultivation medium to each shake flask. Dilution series is prepared by stepwise transfer of 1 mL from flask to flask

• 250 mL shake flasks: Add 18 mL to each shake flask. Dilution series is prepared by stepwise transfer of 2 mL from flask to flask.

Summary

Once you have located the items needed for the experiment proceed to Prepare flasks and media.