# Dilute 1:1 000

Get two dilution vials. Discard 1 mL of diluent of the second dilution vial, so that the second dilution vial is filled to the 9 mL mark.

[dilute-1-100](https://help.sbtinstruments.com/protocols/prepare-your-sample/hit-the-right-concentration/dilute-1-100 "mention") into your first [dilution-vial](https://help.sbtinstruments.com/encyclopedia/item-register/vials-flasks-and-liquids/dilution-vial "mention"), and then [dilute-1-10](https://help.sbtinstruments.com/protocols/prepare-your-sample/hit-the-right-concentration/dilute-1-10 "mention") into your second dilution vial.

 

<figure><img src="https://4216107837-files.gitbook.io/~/files/v0/b/gitbook-x-prod.appspot.com/o/spaces%2FeYTtd7c0A325muFr5lMW%2Fuploads%2Fh5tCcznnRiZqfRh60fj9%2Fdilution%201%201000.png?alt=media&#x26;token=e58f4d4e-81a7-4bd4-b361-e8e473529fd6" alt=""><figcaption></figcaption></figure>

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## Vortex each dilution step thoroughly!

Always vortex thoroughly for at least **10 seconds**. Ensure that a clear cyclone/vortex appears.

You must perform proper vortexing to ensure that the transferred bacteria are well-mixed in the prepared dilution. Without proper vortexing, you may get underestimation of concentration and variation between replicates.
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**Preparing a dilution series with 1:100, 1:1,000 and 1:10,000 dilutions**

The most convenient way of preparing three samples diluted by 1:100, 1:1,000 and 1:10,000 is listed below.

Sample #1 - target dilution 1:100: [dilute-1-100](https://help.sbtinstruments.com/protocols/prepare-your-sample/hit-the-right-concentration/dilute-1-100 "mention") from primary sample.

Sample #2 - target dilution 1:1,000: [dilute-1-10](https://help.sbtinstruments.com/protocols/prepare-your-sample/hit-the-right-concentration/dilute-1-10 "mention") from sample #1. Also, see [dilute-1-1-000](https://help.sbtinstruments.com/protocols/prepare-your-sample/hit-the-right-concentration/dilute-1-1-000 "mention")

Sample #3 - target dilution 1:10,000: [dilute-1-100](https://help.sbtinstruments.com/protocols/prepare-your-sample/hit-the-right-concentration/dilute-1-100 "mention") from sample #1. Also, see [dilute-1-10-000](https://help.sbtinstruments.com/protocols/prepare-your-sample/hit-the-right-concentration/dilute-1-10-000 "mention")
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