> 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/encyclopedia/get-starter-cultures/from-crm-product/inoculate-liquid-culture.md). # Inoculate liquid culture Use the [subcultivation calculator tool](https://www2026.sbtinstruments.com/wp-content/uploads/2025/12/202512-BactoBox-sub-cultivation-calculator.html) to determine the volume needed to seed the growth medium. A typical overnight seeding concentration is 1 x 106 cells/mL. {% hint style="success" %} #### Use Optiseeding workflow to determine suitable seeding concentration The seeding concentration may need adjustment to get a [high-quality culture](/mpd/encyclopedia/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 capture vial in the first field. * Add volume information in the second field. In this case it is 40 mL * Set the desired inoculation concentration in the final field. In this case 1 × 106 cells/mL * Click Calculate V\_add to get the result. {% endstep %} {% step %} **Add starter culture to growth curve flask** Transfer starter culture to growth curve flask and swirl aggressively to disperse the inoculum. 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.

Inoculation of shake flask at well-defined concentration. In this video the 50 mL vial with the red cap represents the capture vial.

{% endstep %} {% endstepper %} ## Summary You have now inoculated the overnight culture at a well-defined starting concentration of 1 x 106 cells/mL. Next step is [Determine concentration after incubation](/mpd/encyclopedia/get-starter-cultures/from-crm-product/determine-concentration-after-incubation.md). --- # Agent Instructions This documentation is published with GitBook. GitBook is the documentation platform designed so that both humans and AI agents can read, navigate, and reason over technical content effectively. Learn more at gitbook.com. ## Querying This Documentation If you need additional information that is not directly available in this page, you can query the documentation by asking a question. Perform an HTTP GET request on the following URL with the `ask` and `goal` query parameters: ``` GET https://help.sbtinstruments.com/mpd/encyclopedia/get-starter-cultures/from-crm-product/inoculate-liquid-culture.md?ask=&goal= ``` `ask` is the immediate question: it should be specific, self-contained, and written in natural language. `goal` is what the user is ultimately trying to achieve, the reason they need the answer. Sharing it helps GitBook give you a better, more relevant answer. A goal is most helpful when it describes the outcome the user wants rather than restating the question. For example, with `ask=how do I create an API token`, a goal like `automate deployments from our CI pipeline` lets GitBook tailor the answer to that use case. The response will contain a direct answer to the question and relevant excerpts and sources from the documentation. Use this mechanism when the answer is not explicitly present in the current page, you need clarification or additional context, or you want to retrieve related documentation sections.