# Get things ready

You will need these items for the experiment. 

<figure><img src="https://4216107837-files.gitbook.io/~/files/v0/b/gitbook-x-prod.appspot.com/o/spaces%2FeYTtd7c0A325muFr5lMW%2Fuploads%2FkThgv75CqTScOu7LjvdX%2FGet_things_ready_cropped.JPG?alt=media&#x26;token=9f90298a-8ad9-4540-af75-4ba8e44f417d" alt=""><figcaption><p>Items required for the experiment: Along with these, ensure to have sterile serological pipettes and a pipette controller available.</p></figcaption></figure>

## Hardware

* [BactoBox® setup](https://help.sbtinstruments.com/encyclopedia/item-register/bactobox-r) including a [flow cell](https://help.sbtinstruments.com/encyclopedia/item-register/consumables/flow-cell) with at least 10 measurements.
* Windows 10/11 computer.
* 10 × shake flasks with vented caps. Preferably [50/125 mL capacity](https://www.dwk.com/na/kimble-kimax-baffled-shake-flask-125-ml-25630-125). Alternative: 2 × [6-deepwell microplates](https://www.enzyscreen.com/product/6-deepwell-plate/).&#x20;
* 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&#x20;

* 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](https://help.sbtinstruments.com/protocols/prepare-your-sample/focus-on-target-objects).&#x20;
* [Dilution vials](https://help.sbtinstruments.com/encyclopedia/item-register/vials-flasks-and-liquids/dilution-vial) either ready-to-use or freshly prepared using the [dilution dispenser](https://help.sbtinstruments.com/encyclopedia/item-register/accessories/dilution-dispenser).
* [Disinfection vials](https://help.sbtinstruments.com/encyclopedia/item-register/vials-flasks-and-liquids/disinfection-vial).

{% hint style="success" %}

## 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.
{% endhint %}

{% hint style="info" icon="flask" %}

## Fill level for different vessels.

* 50/125 mL shake flasks or [6-deepwell microplates](https://www.enzyscreen.com/product/6-deepwell-plate/): 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.&#x20;
  {% endhint %}

## Summary

Once you have located the items needed for the experiment proceed to [prepare-flasks-and-media](https://help.sbtinstruments.com/protocols/optimize-bioprocesses/identify-optimal-inoculum-size/prepare-flasks-and-media "mention").&#x20;