# Inoculate
Use the [subcultivation calculator tool](https://www2026.sbtinstruments.com/wp-content/uploads/2025/12/202512-BactoBox-sub-cultivation-calculator.html) to determine the volume of starter culture needed for the growth curve experiment. Here we are targeting an inoculation concentration of 1 x 107 cells/mL.
{% stepper %}
{% step %}
**Calculate inoculation volume**
1. In Access, triple-click the cell containing Cells/mL x Dil. Press ctrl+c to copy the value.
2. Open a new window and use the [subcultivation calculator tool](https://www2026.sbtinstruments.com/wp-content/uploads/2025/12/202512-BactoBox-sub-cultivation-calculator.html) to determine which volume to add (Tip: To exit full screen mode, press F11 (or Fn+F11 depending on keyboard configuration):
1. Press ctrl+v in the first field to input the concentration from the starter culture shake flask.
2. Add the volume information in the second field.
3. Decide on the target inoculation concentration in the last field.
4. Click Calculate V\_add to get the result.
{% endstep %}
{% step %}
**Add starter culture to growth curve flask**
1. Use proper aseptic technique to transfer starter culture to growth curve flask.
2. Swirl aggressively to disperse the inoculum.
3. Remember to record the inoculation time. Record it on the autoclave tape as a backup, ensuring you have a reference if you forget to document it elsewhere.
{% endstep %}
{% endstepper %}
## Summary
You have now inoculated the growth medium at a well-defined starting concentration of 1 x 107 cells/mL. Next step is to determine the bacterial concentration at T0.
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