# From cryo stock

| Skill level  | Time to complete                                             | Hands-on time                                   | Requirements                                                                                                                                                                                                 |
| ------------ | ------------------------------------------------------------ | ----------------------------------------------- | ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------ |
| Intermediate | <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](/encyclopedia/item-register/bactobox-r/bactobox-r-hardware-changelog.md) <i class="fa-floppy-disk">:floppy-disk:</i> [v2026.02a](/encyclopedia/software.md) |

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

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


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