In this tutorial, we will adapt the workflow built in 10.3 to a PicoGreen Assay.
The ability to accurately quantify the amount of DNA in a sample is required for several applications, such as cDNA library synthesis, cloning, or for the detection of DNA in drug formulations. The PicoGreen Assay is a simple protocol that allows the detection and quantification of double-stranded DNA by using a fluorescent dye that specifically binds to DNA in solution.
From a practical point of view, a PicoGreen protocol is very similar to a Bradford, and as such in this tutorial we will adapt the Bradford workflow (Core elements tutorial 10.3) to create a PicoGreen workflow.
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 copy of the workflow that you created in Core elements tutorial 10.3. To learn how, click here.
Rename the copy Core elements tutorial 10.4. To learn how, click here.
Adapt the workflow to a PicoGreen protocol
To convert the above workflow to a PicoGreen protocol, only two changes are required: modifying the reagent liquid name, and changing the target concentration of the standard dilution curve
In the Define Liquids and Plates element where the Bradford reagent is defined, change it to PicoGreen Reagent.
Set the concentration of the undiluted Standard to 2000 ng/uL.
Modify the dilution settings in the Dilute element (for more information on how to use Dilute, click here). In this case, we want to prepare a linear dilution curve for the standard, ranging from 2-2000 ng/mL. Start by changing the Describe Dilutions parameter to be By Concentration Range.
Specify the Lowest and Highest Concentration values to be 2 and 2000 ng/mL respectively. Untoggle Keep Intermediate Volumes Constant.
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.
Check your work:
To see what your finished workflow to this tutorial should look like simply navigate to the Tutorials and search for Tutorial: 10. Building basic assays.
