5) Setting of Advanced Parameters in combination with Solution Conditions
Click on the Experiment Conditions tab
Enter in the appropriate Solution Conditions (screenshot below for example)
Go back to the Advanced Parameters on the Probe/Primer Design tab and change them if needed
Keep in mind that the delta G contribution of a nearest neighbor basepair decreases with temperature. When increasing the assay temperature either the stem length has to be increased so that the optimal stem dG can be met or the stem dG has to be adjusted (less negative) so that this dG can be met by a stem that has the optimal stem length
