> 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-cryo-stock.md).

# From cryo stock

| Skill level                            | Time to complete                                             | Hands-on time                                   | Requirements                                                                                                                                                                 |
| -------------------------------------- | ------------------------------------------------------------ | ----------------------------------------------- | ---------------------------------------------------------------------------------------------------------------------------------------------------------------------------- |
| :school\_satchel: No skill requirement | <i class="fa-hourglass">:hourglass:</i> 16 hours (*E. coli*) | <i class="fa-stopwatch">:stopwatch:</i> 3 hours | <i class="fa-hard-drive">:hard-drive:</i> [7.6a](/item-register/bactobox-r/bactobox-r-hardware-changelog-retired.md) <i class="fa-floppy-disk">:floppy-disk:</i> ≥ v2026.02a |

To reactivate bacteria from cryo stocks, first add a precise volume of the cryo stock to a shake flask. Incubate the flask to revive the dormant bacteria and allow them to multiply, thereby creating a starter culture of active cells.

{% hint style="info" %}

#### Choose between overnight and same-day reactivation of cells in cryo stocks

<i class="fa-square-1">:square-1:</i> Overnight incubation helps activate cells and prepare starter cultures. We recommend this approach.

<i class="fa-square-2">:square-2:</i> Same-day starter culture is also possible if a higher seeding ratio is used.
{% endhint %}

## <i class="fa-square-1">:square-1:</i> Overnight starter cultures from cryo stocks

[Get starter culture](/mpd/encyclopedia/get-starter-cultures.md) provides a powerful approach to tune the seeding ratio for a [High-quality culture](/mpd/encyclopedia/high-quality-culture.md) when preparing an overnight culture. With this approach it is possible to get extremely consistent starter cultures.

## <i class="fa-square-2">:square-2:</i> Same-day starter culture from cryo stock

Usually it is possible to get a starter culture within the same day if you use a relatively high seeding ratio. The step by step guide is detailed in the subsequent pages.

1. [Get things ready](/mpd/encyclopedia/get-starter-cultures/from-cryo-stock/get-things-ready.md)
2. [Inoculate culture](/mpd/encyclopedia/get-starter-cultures/from-cryo-stock/inoculate-culture.md)
3. [Determine initial concentration](/mpd/encyclopedia/get-starter-cultures/from-cryo-stock/determine-initial-concentration.md)
4. [Determine concentration after incubation](/mpd/encyclopedia/get-starter-cultures/from-cryo-stock/determine-concentration-after-incubation.md)

Proceed to [Get things ready](/mpd/encyclopedia/get-starter-cultures/from-cryo-stock/get-things-ready.md) for initializing the same-day starter culture from cryo stock protocol.


---

# 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-cryo-stock.md?ask=<question>&goal=<user_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.