> For the complete documentation index, see [llms.txt](https://help.sbtinstruments.com/llms.txt). Markdown versions of documentation pages are available by appending `.md` to page URLs; this page is available as [Markdown](https://help.sbtinstruments.com/mpd/workflows/get-starter-cultures/from-liquid-culture/inoculate-culture.md).
# Inoculate culture
Once you know the cell concentration of the inoculum you can calculate the volume necessary to hit a target seeding concentration (here 1 × 107 cells/mL). Use the [subcultivation calculator tool](https://www2026.sbtinstruments.com/wp-content/uploads/2025/12/202512-BactoBox-sub-cultivation-calculator.html) for the calculations.
{% hint style="success" %}
## Use Optiseeding workflow to determine suitable seeding concentration
The seeding concentration may need adjustment to get a [high-quality culture](/mpd/cell-growth/high-quality-culture.md). Do a screening experiment with a broad 1:10 dilution series to find a suitable seeding concentration. See [Optimize starter culture](/mpd/workflows/identify-optimal-inoculum-size.md) for more details.
{% endhint %}
{% stepper %}
{% step %}
**Open the subcultivation calculator**
Open the [subcultivation calculator tool](https://www2026.sbtinstruments.com/wp-content/uploads/2025/12/202512-BactoBox-sub-cultivation-calculator.html) in a browser.
{% endstep %}
{% step %}
**Calculate inoculation volume**
* Fill in cells/mL of the inoculum in the first field.
* Add volume information in the second field. In this case it is 50 mL
* Set the desired inoculation concentration in the final field. In this case 1 × 107 cells/mL
* Click Calculate V\_add to get the result.
{% endstep %}
{% step %}
**Add inoculum culture to growth medium**
Transfer the calculated volume of inoculum to growth curve flask and swirl aggressively to disperse the inoculum.
Record the inoculation time. Note it down on the autoclave tape as a backup, ensuring you have a reference if you forget to document it elsewhere.
Inoculation of shake flask at well-defined concentration. In this video the 50 mL vial with the red cap represents the inoculum.
{% endstep %}
{% endstepper %}
## Summary
Once the growth medium is inoculated, proceed to [Determine initial concentration](/mpd/workflows/get-starter-cultures/from-liquid-culture/determine-initial-concentration.md).
---
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