Most quantification assays run in a lab share the same basic hallmarks; serial dilution of a standard and aliquoting both samples of unknown concentration and the diluted standards to an assay plate. In this tutorial we will guide you through the steps of how to build a basic quantification assay workflow.
This tutorial will build upon the workflow and foundations covered in Core elements tutorial 2.5.
Get started
Read the introduction to this series of tutorials. For more information, click here.
Open the workflow that you created in Core elements tutorial 2.5.
In this workflow we had a set of samples being aliquot in triplicate to an assay plate with a Control being aliquot in triplicate to the same plate.
Add a serial dilution step into the workflow
Let’s modify this workflow to aliquot replicates of a serially diluted standard to our assay plate rather than the control samples that are currently handled.
Create a copy of the workflow that you created in Core elements tutorial 2.5. To learn how, click here.
Rename the copy Core elements tutorial 10.1. To learn how, click here.
Add 1 x Define Liquids And Plates element and 1 x Dilute element to the current workflow.
Delete the connection between the Define Liquids And Plates 2 element and the Aliquot element.
Connect the Liquids output parameter from the Define Liquids And Plates 3 element into the Liquids To Dilute input of the Dilute element.
Connect the Liquids output parameter from the Define Liquids And Plates 2 element into the Diluents input of the Dilute element.
Connect the Dilutions output parameter from the Dilute 1 element into the Liquids To Aliquot input of the Aliquot 2 element.
Specify your dilution parameters
Define your standard in the Define Liquids And Plates 3 element.
Define your diluent in the Define Liquids And Plates 2 element.
Define your dilution parameters, here the Dilution Location is set to New Location, Dilution Settings is set to Serial, and Describe Dilutions is set to By Factor.
Specify the diluent you want to use for all dilutions.
Specify dilution options.
Specify Layout Options
Update the plate layout
Click on the Describe Layouts 1 element to open the parameters panel and click on the EDIT PLATE CONTENTS button.
Delete the two well sets that are already defined there by clicking on the trash can icon to the right hand side of them.
Specify two new well sets, one for the samples and one for the diluted standards.
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.
Notice here that Synthace now makes dilutions of BSA to an intermediate Dilution Plate then aliquots samples and diluted standards in triplicate to the single Assay Plate that you defined in the Describe Layouts element.