# 1:100 dilutions

For every 30 minutes, collect a fresh sample and repeat the procedure from the 1:100 dilution.

{% hint style="success" %}

#### <i class="fa-spray-can">:spray-can:</i> Do a *Clean* program after each measurement.

{% endhint %}

{% hint style="success" %}

#### <i class="fa-hourglass">:hourglass:</i> Start a 30-minute countdown to remind you to collect the next sample.

{% endhint %}

{% stepper %}
{% step %}
**Prepare the sample for BactoBox® measurement**

1. Use a serological pipette to transfer a sample from the shake flask to a vial.
2. Use a permanent marker to label the vial cap with the sampling time.
3. Start a 30-minute countdown to remind you to collect the next sample.
4. Vortex 30 seconds at max speed to disaggregate bacteria.
5. Make a 1:100 dilution:
   1. Transfer 101 µL of your sample to 10 mL of diluent.
   2. Vortex 10 seconds at max speed.
6. Transfer the tubing kit to the diluted sample.

<figure><img src="/files/tUo0C4h2nhT5bte8twIs" alt=""><figcaption></figcaption></figure>
{% endstep %}

{% step %}
**Measure the diluted sample on BactoBox®**

1. Click the <kbd><mark style="background-color:purple;">Measure<mark style="background-color:purple;"></kbd> button in Access.
2. Add metadata:
   1. Dilution: <kbd>100</kbd>.
   2. Label: <kbd>E\_coli\_growth</kbd>.
      {% endstep %}

{% step %}
**Clean the device**

1. Transfer the tubing kit to the disinfection vial.
2. Click <kbd>Clean</kbd>.
   {% endstep %}
   {% endstepper %}

## Summary

The 1:100 dilution scheme works well for the measurements during lag and early-exponential growth stage. As the concentration increases, you'll need to dilute more. We will explain this in detail in the next section.


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