Use 1:100 dilutions in lag and early-exponential growth stage

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

Do a Clean program after each measurement.

Start a 30-minute countdown to remind you to collect the next sample.

Prepare the T0 sample for BactoBox® measurement

1. Use a serological pipette to collect a sample from the shake flask.

2. Return the shake flask to the incubator shaker.

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.

Measure the diluted samples in Access

1. Click the Measure button in Access.

2. Add metadata:

   1. Dilution: 100.

   2. Label: E_coli_growth.

Clean the device

1. Transfer the tubing kit to the disinfection vial.

2. Click Clean.

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.