# Rehydrate and streak

The first goal to obtain a streaked agar plate of single, well-isolated colonies. The [lyfo disk<sup>TM</sup>](https://www.microbiologics.com/item-type/Product/product-format/LYFO-DISK) is used as a starting material.

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#### See [Microbiologics](https://www.microbiologics.com/item-type/Product/product-format/Epower) [lyfo disk<sup>TM</sup>](https://www.microbiologics.com/item-type/Product/product-format/LYFO-DISK) documentation for full details

We use an adapted version of the [lyfo disk<sup>TM</sup>](https://www.microbiologics.com/item-type/Product/product-format/LYFO-DISK) rehydration protocol. For full description please inspect the [supporting material provided by Microbiologics](https://www.microbiologics.com/item-type/Product/product-format/LYFO-DISK).
{% endhint %}

## Step by step

{% stepper %}
{% step %}
**Transfer disk to capture tube**

Use aseptic technique to transfer a LYFO DISK to a preheated (37 °C) 5 mL capture tube containing glass beads.
{% endstep %}

{% step %}
**Dissolve disk and disperse cells**

Rehydrate the disk by bead-beating, manual shaking or vortexing for 1 min at max speed (or until the suspension is homogenous)
{% endstep %}

{% step %}
**Streak agar plate**

Dip a sterile 1 µL inoculate loop into the suspension and [streak](https://microbenotes.com/streak-plate-method-principle-methods-significance-limitations/) to obtain single, well-resolved colonies.
{% endstep %}

{% step %}
**Incubate**

Incubate at optimal growth conditions until colonies appear (typically overnight). Avoid over-incubation as this may lead to presence of dead cells.
{% endstep %}
{% endstepper %}

## Summary

The [Lyfo disk<sup>TM</sup>](https://www.microbiologics.com/item-type/Product/product-format/LYFO-DISK) is rehydrated and an agar plate is streaked to get single colonies. Next step is to [Collect and disperse](/mpd/workflows/get-starter-cultures/from-crm-product/collect-and-disperse.md).


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