Core elements tutorial 3.4
Here we will cover the process of performing a direct dilution on a defined liquid to a specific set of concentrations. This dilution process will prepare a set of diluted liquids, but is focussed on doing this with a dispenser, where true serial dilution is not possible.
Get started
Read the introduction to this series of tutorials, then complete all of the previous tutorials in the series. For more information, click here.
Create a workflow in the builder. To learn how, click here.
Rename the workflow Core elements tutorial 3.4. To learn how, click here.
Select the device on which you want to run your workflow. To learn how, click here.
Select a default plate type. To learn how, click here.
Define liquids
First, you must define the liquids required for the dilution process. These are i) the liquids you intend to dilute, ii) a diluent. For detailed instructions on defining liquids see here.
Open the Elements menu and search for the element called Define Liquids And Plates and click on the element tile to add an instance of this element to your canvas.
Click on the element to highlight it. This will open the parameters panel for this specific element instance.
In the parameters panel select Table Editor in the Define From parameter.
Click on Add Spreadsheet button to open the table editor.
You can define liquids by filling in the table. Here we have defined Tris buffer and Water in Liquid Names. We have also defined Concentrations and Concentration Units. The Tris buffer has been tagged with the Tag
Bufferand Water has been tagged withDiluent. Once you have defined the liquids, click Done in the bottom right-hand corner.
For clarity re-name the Define Liquids and Plates element to Define Buffer in the Instance Name field.
In this workflow, the volume and plate type are not in the Liquid Definition and will automatically be assigned based on the workflow requirements.
Repeat this process to define the liquid you intend to use as a diluent.
Rename the second instance of the Define Liquids and Plates to Define Diluent.
Dilute the Liquids
Specify dilution behaviour
Add the Dilute element to the workflow builder from the Elements menu.
Click on the Dilute element to display the element parameters for this instance.
Under Global Options specify the Dilution Location as New Location.
Set the Dilution Mode to Direct.
Set Describe Dilutions to By Concentration Range.
Provide liquids to dilute and specify dilution location
We have now established how we would like to carry out the dilutions in Synthace. Now we will define which liquids we would like to dilute, what we will dilute them with and buy how much.
Connect the Liquids output from Define Buffer to Liquids to Dilute.
Connect the Liquids output from Define Diluent to Diluent.
Specify the Diluent To Use Per Liquid parameter. In this case, we want to use the Water we defined in Define Diluent for all our substrates.
Note: if you wish to override the liquid policy of the diluted liquids you can do so here.
Define the Target Final Volume.
Define the Lowest Concentration and Highest Concentration for the dilution series and the number of Replicates and Dilution Number.
Note: if you wish to override the liquid policy of the diluted liquids you can do so here:
Specifying dilution layout and output plate type
With the dilution behaviour defined we now need to specify where the dilutions will take place.
Select Let Synthace Optimize for Layout and Replicate Grouping.
Define a Plate Name. This can be any alphanumeric name such as
dilution_plate.Select a Plate Type from your plate library. Take care to ensure the plate type you select can accommodate the target volume for your dilution.
Note: For more details of using layouts in your workflows head to this tutorial.
Simulate the workflow
Check that the device that you selected can follow the instructions that you prepared. To learn how, click here.
Preview the execution
After you simulate the workflow, click View Simulation to open the simulation details.
Open the Preview tab, then click through the steps to check that the instructions that Synthace has generated are correct.
You will see the input plate containing the buffer and your diluent. These will then be dispensed to the output plate, first the diluent and then the buffer to yield liquids with the specified buffer concentration, in triplicate with a final volume of 100 ul.
Check your work:
To see what your finished workflow to this tutorial should look like simply navigate to the Tutorials and search for Tutorial 3: Dilute.
Challenges
Repeat this process for a second buffer. Layout the dilution series in the plate by column, with each replicate in a separate column. Leave a blank row between the sets of dilutions for each buffer. See the tutorial here for tips on how to do this.