# From agar plate colony

| Skill level  | Time to complete                                                | Hands-on time                                   | Requirements                                                                                                                                                                                                 |
| ------------ | --------------------------------------------------------------- | ----------------------------------------------- | ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------ |
| Intermediate | <i class="fa-hourglass">:hourglass:</i> 16-24 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) |

A fresh colony from an agar plate is a good starting point to ensure a single-species (axenic) culture. The standard workflow uses overnight incubation of the liquid culture, but it is often possible to get a same-day starter culture from an agar plate (see hint box).

<figure><img src="/files/mNhgQwJGKWRER6VElGy9" alt=""><figcaption><p>Illustration of workflow. A fresh colony is collected and transferrred to a capture tube. Cell clumps are disaggregated using bead-beater or vortex mixer. Finally, a sample of capture tube is diluted and subjected to BactoBox® measurement.</p></figcaption></figure>

{% hint style="info" icon="sneaker-running" %}

## Get a same-day starter culture

* If you want a same-day starter culture, try using higher seeding concentration, e.g. 5× 10<sup>7</sup> cells/mL.
* Check the liquid culture frequently for increase in cell concentration, e.g. for each 2 hours.
* We recommend that - prior to use - the active starter culture should have cell concentration is at least tenfold higher than at T<sub>0</sub>.
  {% endhint %}

The protocol assumes that you have already used the [streak plate method](https://microbenotes.com/streak-plate-method-principle-methods-significance-limitations/) to get fresh colonies on an agar plate. The steps are outlined below.

1. [Get things ready](/mpd/workflows/get-starter-cultures/from-agar-plate-colony/get-things-ready.md)
2. [Collect and disperse bacteria from agar plate](/mpd/workflows/get-starter-cultures/from-agar-plate-colony/collect-and-disperse.md)
3. [Determine cells/mL in harvest tube](/mpd/workflows/get-starter-cultures/from-agar-plate-colony/determine-concentration.md)
4. [Inoculate liquid culture](/mpd/workflows/get-starter-cultures/from-agar-plate-colony/inoculate-liquid-culture.md)
5. [Determine concentration of overnight culture](/mpd/workflows/get-starter-cultures/from-agar-plate-colony/determine-concentration-after-incubation.md)

Next, the [get things ready](/mpd/workflows/get-starter-cultures/from-agar-plate-colony/get-things-ready.md) section will provide you with a check-list of necessary items.


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