If you have a set of samples of varying concentration and you want to prepare new samples from these all at the same concentration, try using the Normalise element. We will explain how to normalize the concentration of a set of liquids using the Normalize element. The normalized samples will be prepared in a new output plate.
In this example we will normalize the concentration of a set of previously purified plasmid DNA. The normalized samples will be prepared in a new output plate, thus maintaining the concentration of the original plasmid stocks.
Get started
Read the introduction to this series of tutorials. For more information, click here.
Create a workflow in the builder. To learn how, click here.
Rename the workflow Core elements tutorial 4.1. 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
In the Workflow Builder add three elements to your canvas: 2 x Define Liquids And Plates and 1 x Normalize. Rename the liquid definition elements to Define Plasmid DNA and Define Diluent.
Connect the Output Liquids parameter from Define Plasmid DNA into the Liquids To Normalize input parameter of Normalize.
Connect the output Liquids parameter from Define Diluent element into the Diluents input of Normalize.
Define your samples that are intended for normalization. For more information on how, see here. In this example we have defined 16 plasmid dna samples of varying concentrations.
Define your Diluent(s) in the Define Diluent element.
Note: Defining only one or two liquids, like diluents, is often easiest using the Parameters option in the Define Liquids And Plates element.
Describe normalization behaviour
In the Normalize element select the New Plate option in the Normalization Location parameter and the By Concentration option in the Normalization Mode parameter.
Specify which diluent you want to use for each of your normalizations in the Diluent To Use Per Liquid parameter.
In the Normalization Options group of the Normalize element parameters specify the Target Final Volume and Target Concentration
Specify the number of Replicates of the normalized samples you want to prepare.
In the Layout Options of the Normalize element parameters specify how you want your replicates grouped, for this example we will use Let Synthace Optimize.
Provide a plate name in Plate Set Names and define the Plate Type that the normalized samples will be prepared in. Ensure that the well size of this plate is sufficient for the Target Final Volume you have specified.
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 that different volumes of diluent are aliquoted into a new plate and the samples are then transferred into these wells, with replicates adjacent to each other. The final concentration of resulting liquids is the same across all the wells.
Check your work:
To see what your finished workflow to this tutorial should look like simply navigate to the Tutorials and search for Tutorial: 4. Normalize.
Challenges
Here we have normalized the samples according to their concentration. It is also possible to normalize based on the concentration of a specific sub component. See if you can modify the workflow to normalize a set of sample by a specific subcomponent.
Hint: Subcomponent Concentrations can be added in the Define Liquids and Plates element.