# Dilute 1:100
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## Dilute 1:100 is your go-to dilution method
It is both easy and fast. Usually, you want to start with a 1:10 000 dilution first, which is just *two* 1:100 dilutions in a row.
[**Measure the most diluted sample first**](#measure-the-most-diluted-sample-first).
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## By the book
The “by the book” method is to prepare aliquots of 9900 µL diluent and add 100 µL of sample. This works if you have the patience and a bottle-top dispenser or a P10,000 pipette.
## Our shortcut
There is a much easier way forward: Add 101 µL of your culture to the 10 mL in a [dilution vial](https://help.sbtinstruments.com/encyclopedia/item-register/vials-flasks-and-liquids/dilution-vial). This gives you a 1:100.01 dilution, which is *very* close to the target of 1:100. Once mixed together, we call it your *sample vial* from now on.
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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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## Your sample vial is for *single use* only
Once you run the liquid through BactoBox®, the properties change *significantly*.
**Always discard your sample vial after use!**
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## Need to dilute 1:10 000? Or 1:1 000 000?
Simply repeat the process described above. A 1:100 dilution followed by another 1:100 dilution gives a 1:10 000 dilution. Add another 1:100 dilution and you get a total dilution of 1:1 000 000.
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For every single 1:100 dilution you do, add two zeroes to the overall dilution factor.
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## Measure the most diluted sample first
If you don't know the concentration in your sample vial, it is best to play it safe. Prepare a dilution series as pictured above. First, measure on the most diluted sample. Then, move towards less diluted samples step by step. Stop when the concentration is within 0.5–4 million cells/mL.
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## Target 0.5–4 million cells/mL for fast results
The more things to count, the faster we are certain about the concentration. Give BactoBox® something to work with and it triggers the *Fast results* feature :fast\_forward:.
Make a dilution series and [**measure the most diluted sample first**](#measure-the-most-diluted-sample-first). Aim for 0.5–4 million cells/mL.
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## Is the final dilution factor in your sample vial 1:100?
You might run into conductivity-related errors. Avoid these with a [custom diluent](https://help.sbtinstruments.com/protocols/hit-the-right-conductivity#i-dilute-1-100-or-thereabout).
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